LOCUS       SEGA092                6539 bp ds-DNA     linear   UNA 09-SEP-2021
DEFINITION  natural linear DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6539)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Sep 9, 2021 from SnapGene 4.2.11
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     CDS             1..528
                     /codon_start=1
                     /label=glmS
                     /note="glmS"
                     /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI
                     ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR
                     ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD
                     QPRNLAKSVTVE"
     terminator      538..594
                     /label=L3S2P11 Terminator
                     /note="L3S2P11 Terminator"
     CDS             complement(605..1753)
                     /codon_start=1
                     /gene="galK"
                     /gene_synonym="ECK0746; galA; JW0740"
                     /locus_tag="b0757"
                     /product="galactokinase"
                     /function="enzyme; Degradation of small molecules: Carbon 
                     compounds"
                     /EC_number="2.7.1.6"
                     /label=galK
                     /note="galK"
                     /db_xref="GI:16128725"
                     /db_xref="ASAP:ABE-0002568"
                     /db_xref="UniProtKB/Swiss-Prot:P0A6T3"
                     /db_xref="EcoGene:EG10363"
                     /db_xref="GeneID:945358"
                     /protein_id="NP_415278.1"
                     /translation="MSLKEKTQSLFANAFGYPATHTIQAPGRVNLIGEHTDYNDGFVLP
                     CAIDYQTVISCAPRDDRKVRVMAADYENQLDEFSLDAPIVAHENYQWANYVRGVVKHLQ
                     LRNNSFGGVDMVISGNVPQGAGLSSSASLEVAVGTVLQQLYHLPLDGAQIALNGQEAEN
                     QFVGCNCGIMDQLISALGKKDHALLIDCRSLGTKAVSMPKGVAVVIINSNFKRTLVGSE
                     YNTRREQCETGARFFQQPALRDVTIEEFNAVAHELDPIVAKRVRHILTENARTVEAASA
                     LEQGDLKRMGELMAESHASMRDDFEITVPQIDTLVEIVKAVIGDKGGVRMTGGGFGGCI
                     VALIPEELVPAVQQAVAEQYEAKTGIKETFYVCKPSQGAGQC"
     promoter        complement(1783..1830)
                     /label=EM7 promoter
                     /note="EM7 promoter"
                     /note="synthetic bacterial promoter"
     terminator      1843..1886
                     /label=Terminator rrnB T1
                     /note="Terminator rrnB T1"
     CDS             complement(1899..2744)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="rhaR"
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(2818..3654)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter"
                     /label=rhaS
                     /note="rhaS"
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        3800..3918
                     /label=rhaB promoter
                     /note="rhaB promoter"
                     /note="promoter of the E. coli rhaBAD operon, conferring 
                     tight induction with L-rhamnose and repression with 
                     D-glucose in the presence of RhaR and RhaS (Giacalone et 
                     al., 2006)"
     RBS             3930..3956
                     /note="TIR"
                     /note="high TIR"
     CDS             3948..4013
                     /codon_start=1
                     /gene="pelB (fragment)"
                     /product="leader peptide for secretion"
                     /label=pelB signal sequence
                     /note="pelB signal sequence"
                     /translation="MKYLLPTAAAGLLLLAAQPAMA"
     CDS             4014..4724
                     /codon_start=1
                     /product="superfolder GFP"
                     /label=sfGFP
                     /note="sfGFP"
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      4728..4776
                     /label=L3S3P22 Terminator (synthetic)
                     /note="L3S3P22 Terminator"
     promoter        4787..4815
                     /label=Promoter 3BCD2
                     /note="Promoter 3BCD2"
     CDS             4855..6045
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="TcR"
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMIGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     terminator      6057..6097
                     /label=rrnC
                     /note="rrnC"
     CDS             6292..6539
                     /codon_start=1
                     /label=pstS
                     /note="pstS"
                     /translation="MKVMRTTVATVVAATLSMSAFSVFAEASLTGAGATFPAPVYAKWA
                     DTYQKETGNKVNYQGIGSSGGVKQIIANTVDFGASDA"
ORIGIN
        1 gtgcatggtc tgcaggcgct gccgagccgt attgagcaga tgctgtctca ggacaaacgc
       61 attgaagcgc tggcagaaga tttctctgac aaacatcacg cgctgttcct gggccgtggc
      121 gatcagtacc caatcgcgct ggaaggcgca ttgaagttga aagagatctc ttacattcac
      181 gctgaagcct acgctgctgg cgaactgaaa cacggtccgc tggcgctaat tgatgccgat
      241 atgccggtta ttgttgttgc accgaacaac gaattgctgg aaaaactgaa atccaacatt
      301 gaagaagttc gcgcgcgtgg cggtcagttg tatgtcttcg ccgatcagga tgcgggtttt
      361 gtaagtagcg ataacatgca catcatcgag atgccgcatg tggaagaggt gattgcaccg
      421 atcttctaca ccgttccgct gcagctgctg gcttaccatg tcgcgctgat caaaggcacc
      481 gacgttgacc agccgcgtaa cctggcaaaa tcggttacgg ttgagtaata aatggatctc
      541 ggtaccaaat tccagaaaag agacgctttc gagcgtcttt tttcgttttg gtccaccgta
      601 ttcatcagca ctgtcctgct ccttgtgatg gtttacaaac gtaaaaagtc tctttaatac
      661 ctgtttttgc ttcatattgt tcagcgacag cttgctgtac ggcaggcacc agctcttccg
      721 ggatcagcgc gacgatacag ccgccaaatc cgccgccggt catgcgtacg ccacctttgt
      781 cgccaatcac agctttgacg atttctacca gagtgtcaat ttgcggcacg gtgatttcga
      841 aatcatcgcg catagaggca tgagactccg ccatcaactc gcccatacgt ttcaggtcgc
      901 cttgctccag cgcgctggca gcttcaacgg tgcgggcgtt ttcagtcagt atatgacgca
      961 cgcgttttgc cacgatcggg tccagttcat gcgcaacagc gttgaactct tcaatggtga
     1021 catcacgcag ggctggctgc tggaagaaac gcgcaccggt ttcgcactgt tcacgacggg
     1081 tgttgtattc gctgccaacc agggtacgtt tgaagttact gttgatgatg acgacagcca
     1141 cacctttggg catggaaact gctttggtcc ccagtgagcg gcaatcgatc agcaaggcat
     1201 gatctttctt gccgagcgcg gaaattagct gatccatgat cccgcagtta cagcctacaa
     1261 actggttttc tgcttcctga ccgttaagcg cgatttgtgc gccgtccagc ggcagatgat
     1321 aaagctgctg caatacggtt ccgaccgcga cttccagtga agcggaagaa cttaacccgg
     1381 caccctgcgg cacattgccg ctgatcacca tgtccacgcc gccgaagctg ttgttacgca
     1441 gttgcagatg tttcaccacg ccacgaacgt agttagccca ttgatagttt tcatgtgcga
     1501 caatgggcgc atcgagggaa aactcgtcga gctgattttc ataatcggct gccatcacgc
     1561 gaactttacg gtcatcgcgt ggtgcacaac tgatcacggt ttgataatca atcgcgcagg
     1621 gcagaacgaa accgtcgttg tagtcggtgt gttcaccaat caaattcacg cggccaggcg
     1681 cctgaatggt gtgagtggca gggtagccaa atgcgttggc aaacagagat tgtgtttttt
     1741 ctttcagact catgattgcc tcctgggtta gttcctcacc ttgtcgtatt atactatgcc
     1801 gatatactat gccgatgatt aattgtcaac ctttaatgaa ttataaaacg aaaggctcag
     1861 tcgaaagact gggcctttcg ttttatgcgt tgcgcttaat ctttctgcga attgagatga
     1921 cgccactggc tgggcgtcat cccggtttcc cgggtaaaca ccaccgaaaa atagttacta
     1981 tcttcaaagc cacattcggt cgaaatatca ctgattaaca ggcggctatg ctggagaaga
     2041 tattgcgcat gacacactct gacctgtcgc agatattgat tgatggtcat tccagtctgc
     2101 tggcgaaatt gctgacgcaa aacgcgctca ctgcacgatg cctcatcaca aaatttatcc
     2161 agcgcaaagg gacttttcag gctagccgcc agccgggtaa tcagcttatc cagcaacgtt
     2221 tcgctggatg ttggcggcaa cgaatcactg gtgtaacgat ggcgattcag caacatcacc
     2281 aactgcccga acagcaactc agccatttcg ttagcaaacg gcacatgctg actactttca
     2341 tgctcaagct gaccaataac ctgccgcgcc tgcgccatcc ccatgctacc taagcgccag
     2401 tgtggttgcc ctgcgctggc gttaaatccc ggaatcgccc cctgccagtc aagattcagc
     2461 ttcagacgct ccgggcaata aataatattc tgcaaaacca gatcgttaac ggaagcgtag
     2521 gagtgtttat catcagcatg aatgtaaaag agatcgccac gggtaatgcg ataagggcga
     2581 tcgttgagta catgcaggcc attaccgcgc cagacaatca ccagctcaca aaaatcatgt
     2641 gtatgttcag caaagacatc ttgcggataa cggtcagcca cagcgactgc ctgctggtcg
     2701 ctggcaaaaa aatcatcttt gagaagtttt aactgatgcg ccaccgtggc tacctcggcc
     2761 agagaacgaa gttgattatt cgcaatatgg cgtacaaata cgttgagaag attcgcgtta
     2821 ttgcagaaag ccatcccgtc cctggcgaat atcacgcggt gaccagttaa actctcggcg
     2881 aaaaagcgtc gaaaagtggt tactgtcgct gaatccacag cgataggcga tgtcagtaac
     2941 gctggcctcg ctgtggcgta gcagatgtcg ggctttcatc agtcgcaggc ggttcaggta
     3001 tcgctgaggc gtcagtcccg tttgctgctt aagctgccga tgtagcgtac gcagtgaaag
     3061 agaaaattga tccgccacgg catcccaatt cacctcatcg gcaaaatggt cctccagcca
     3121 ggccagaagc aagttgagac gtgatgcgct gttttccagg ttctcctgca aactgctttt
     3181 acgcagcaag agcagtaatt gcataaacaa gatctcgcga ctggcggtcg agggtaaatc
     3241 attttcccct tcctgctgtt ccatctgtgc aaccagctgt cgcacctgct gcaatacgct
     3301 gtggttaacg cgccagtgag acggatactg cccatccagc tcttgtggca gcaactgatt
     3361 cagcccggcg agaaactgaa atcgatccgg cgagcgatac agcacattgg tcagacacag
     3421 attatcggta tgttcataca gatgccgatc atgatcgcgt acgaaacaga ccgtgccacc
     3481 ggtgatggta tagggctgcc cattaaacac atgaataccc gtgccatgtt cgacaatcac
     3541 aatttcatga aaatcatgat gatgttcagg aaaatccgcc tgcgggagcc ggggttctat
     3601 cgccacggac gcgttaccag acggaaaaaa atccacacta tgtaatacgg tcatactggc
     3661 ctcctgatgt cgtcaacacg gcgaaatagt aatcacgagg tcaggttctt accttaaatt
     3721 ttcgacggaa aaccacgtaa aaaacgtcga tttttcaaga tacagcgtga attttcagga
     3781 aatgcggtga gcatcacatc accacaattc agcaaattgt gaacatcatc acgttcatct
     3841 ttccctggtt gccaatggcc cattttcttg tcagtaacga gaaggtcgcg aatccaggcg
     3901 ctttttagac tggtcgtata ctagagatta aagaggagaa attaattatg aaatatctgc
     3961 tgccgaccgc tgctgctggt ctgctgctcc tcgctgccca gccggcgatg gcccgtaaag
     4021 gcgaagagct gttcactggt gtcgtcccta ttctggtgga actggatggt gatgtcaacg
     4081 gtcataagtt ttccgtgcgt ggcgagggtg aaggtgacgc aactaatggt aaactgacgc
     4141 tgaagttcat ctgtactact ggtaaactgc cggtaccttg gccgactctg gtaacgacgc
     4201 tgacttatgg tgttcagtgc tttgctcgtt atccggacca catgaagcag catgacttct
     4261 tcaagtccgc catgccggaa ggctatgtgc aggaacgcac gatttccttt aaggatgacg
     4321 gcacgtacaa aacgcgtgcg gaagtgaaat ttgaaggcga taccctggta aaccgcattg
     4381 agctgaaagg cattgacttt aaagaagacg gcaatatcct gggccataag ctggaataca
     4441 attttaacag ccacaatgtt tacatcaccg ccgataaaca aaaaaatggc attaaagcga
     4501 attttaaaat tcgccacaac gtggaggatg gcagcgtgca gctggctgat cactaccagc
     4561 aaaacactcc aatcggtgat ggtcctgttc tgctgccaga caatcactat ctgagcacgc
     4621 aaagcgttct gtctaaagat ccgaacgaga aacgcgatca catggttctg ctggagttcg
     4681 taaccgcagc gggcatcacg catggtatgg atgaactgta caaatgacca attattgaag
     4741 gccgctaacg cggccttttt ttgtttctgg tctcccaaaa atttatttgc ttattaatca
     4801 tccggctcgt ataatgtgtg gagggcccaa gttcacttaa aaaggagaat agttatgaaa
     4861 tcgaacaatg cgctcatcgt catcctcggc accgtcaccc tggatgctgt aggcataggc
     4921 ttggttatgc cggtactgcc gggcctcttg cgggatatcg tccattccga cagcatcgcc
     4981 agtcactatg gcgtgctgct agcgctatat gcgttgatgc aatttctatg cgcacccgtt
     5041 ctcggagcac tgtccgaccg ctttggccgc cgcccagtcc tgctcgcttc gctacttgga
     5101 gccactatcg actacgcgat catggcgacc acacccgtcc tgtggattct ctacgccgga
     5161 cgcatcgtgg ccggcatcac cggcgccaca ggtgcggttg ctggcgccta tatcgccgac
     5221 atcaccgatg gggaagatcg ggctcgccac ttcgggctca tgagcgcttg tttcggcgtg
     5281 ggtatggtgg caggccccgt ggccggggga ctgttgggcg ccatctccct gcacgcacca
     5341 ttccttgcgg cggcggtgct caacggcctc aacctactac tgggctgctt cctaatgcag
     5401 gagtcgcata agggagagcg ccgtccgatg cccttgagag ccttcaaccc agtcagctcc
     5461 ttccggtggg cgcggggcat gaccattgtg gccgcactta tgactgtctt ctttatcatg
     5521 caactcgtag gacaggtgcc ggcagcgctc tgggtcattt tcggcgagga ccgctttcgc
     5581 tggagcgcga cgatgatcgg cctgtcgctt gcggtattcg gaatcttgca cgccctcgct
     5641 caagccttcg tcactggtcc cgccaccaaa cgtttcggcg agaagcaggc cattatcgcc
     5701 ggcatggcgg ccgacgcgct gggctacgtc ttgctggcgt tcgcgacgcg aggctggatg
     5761 gccttcccca ttatgattct tctcgcttcc ggcggcatcg ggatgcccgc gttgcaggcc
     5821 atgctgtcca ggcaggtaga tgacgaccat cagggacagc ttcaaggatc gctcgcggct
     5881 cttaccagcc taacttcgat cactggaccg ctgatcgtca cggcgattta tgccgcctcg
     5941 gcgagcacat ggaacgggtt ggcatggatt gtaggcgccg ccctatacct tgtctgcctc
     6001 cccgcgttgc gtcgcggtgc atggagccgg gccacctcga cctgaggaca attgtccaga
     6061 taaaaaaaat ccttagcttt cgctaaggat gatttctaaa atttttctgt catctcttcg
     6121 ttattaatgt ttgtaattga ctgaatatca acgcttattt aaatcagact gaagacttta
     6181 tctctctgtc ataaaactgt catattcctt acatataact gtcacctgtt tgtcctattt
     6241 tgcttctcgt agccaacaaa caatgcttta tgaatcctcc caggagacat tatgaaagtt
     6301 atgcgtacca ccgtcgcaac tgttgtcgcc gcgaccttat cgatgagtgc tttctctgtg
     6361 tttgcagaag caagcctgac aggtgcaggt gcaaccttcc ctgcgccggt gtatgccaaa
     6421 tgggctgaca cttaccagaa agaaaccggt aataaagtta actaccaggg tatcggttct
     6481 tccggtggcg taaaacagat tatcgctaat accgttgatt ttggtgcctc tgacgcgcc
//
