LOCUS       SEGA087                6529 bp ds-DNA     linear   UNA 09-SEP-2021
DEFINITION  LMBP 12923.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6529)
  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="low TIR"
     CDS             3948..3968
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /note="TEV site"
                     /translation="MENLYFQ"
     misc_feature    3969..4004
                     /label=PROTi-Linker
                     /note="PROTi-Linker"
     CDS             4005..4715
                     /codon_start=1
                     /product="green fluorescent protein"
                     /label=GFP
                     /note="GFP"
                     /translation="SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDGN
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKAN
                     FKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     terminator      4719..4767
                     /label=L3S3P22 Terminator (synthetic)
                     /note="L3S3P22 Terminator"
     promoter        4778..4806
                     /label=Promoter 3BCD2
                     /note="Promoter 3BCD2"
     CDS             4846..6036
                     /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      6047..6087
                     /label=rrnC
                     /note="rrnC"
     CDS             6282..6529
                     /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 acgggggatg gagaacctgt
     3961 acttccagtt tcgtagcaaa ggagaagaac ttgtcactgg agttagcaaa ggagaagaac
     4021 ttttcactgg agttgtgcca attcttgttg aattagatgg tgatgttaat gggcacaaat
     4081 tttctgtcag tggagagggt gaaggtgatg ctacatacgg aaaactcacc cttaaattta
     4141 tttgcactac tggaaaacta cctgttccat ggccaacact tgtcactact ctgacctatg
     4201 gcgttcaatg cttttcccgt tatccggatc acatgaaacg gcatgacttt ttcaagagtg
     4261 ccatgcccga aggttatgta caggaacgca ctatatcttt caaagatgac gggaactaca
     4321 agacgcgtgc tgaagtcaag tttgaaggtg atacccttgt taatcgtatc gagttaaagg
     4381 gtattgattt taaagaagat ggaaacattc tcggacacaa actagagtac aactataact
     4441 cacacaatgt atacatcacg gcagacaaac aaaagaatgg aatcaaagct aacttcaaaa
     4501 ttcgccacaa cattgaagat ggttccgttc aactagcaga ccattatcaa caaaatactc
     4561 caattggcga tggccctgtc cttttaccag acaaccatta cctgtcgaca caatctgccc
     4621 tttcgaaaga tcccaacgaa aagcgtgacc acatggtcct tcttgagttt gtaactgctg
     4681 ctgggattac acatggcatg gatgagctct acaaatgacc aattattgaa ggccgctaac
     4741 gcggcctttt tttgtttctg gtctcccaaa aatttatttg cttattaatc atccggctcg
     4801 tataatgtgt ggagggccca agttcactta aaaaggagaa tagttatgaa atcgaacaat
     4861 gcgctcatcg tcatcctcgg caccgtcacc ctggatgctg taggcatagg cttggttatg
     4921 ccggtactgc cgggcctctt gcgggatatc gtccattccg acagcatcgc cagtcactat
     4981 ggcgtgctgc tagcgctata tgcgttgatg caatttctat gcgcacccgt tctcggagca
     5041 ctgtccgacc gctttggccg ccgcccagtc ctgctcgctt cgctacttgg agccactatc
     5101 gactacgcga tcatggcgac cacacccgtc ctgtggattc tctacgccgg acgcatcgtg
     5161 gccggcatca ccggcgccac aggtgcggtt gctggcgcct atatcgccga catcaccgat
     5221 ggggaagatc gggctcgcca cttcgggctc atgagcgctt gtttcggcgt gggtatggtg
     5281 gcaggccccg tggccggggg actgttgggc gccatctccc tgcacgcacc attccttgcg
     5341 gcggcggtgc tcaacggcct caacctacta ctgggctgct tcctaatgca ggagtcgcat
     5401 aagggagagc gccgtccgat gcccttgaga gccttcaacc cagtcagctc cttccggtgg
     5461 gcgcggggca tgaccattgt ggccgcactt atgactgtct tctttatcat gcaactcgta
     5521 ggacaggtgc cggcagcgct ctgggtcatt ttcggcgagg accgctttcg ctggagcgcg
     5581 acgatgatcg gcctgtcgct tgcggtattc ggaatcttgc acgccctcgc tcaagccttc
     5641 gtcactggtc ccgccaccaa acgtttcggc gagaagcagg ccattatcgc cggcatggcg
     5701 gccgacgcgc tgggctacgt cttgctggcg ttcgcgacgc gaggctggat ggccttcccc
     5761 attatgattc ttctcgcttc cggcggcatc gggatgcccg cgttgcaggc catgctgtcc
     5821 aggcaggtag atgacgacca tcagggacag cttcaaggat cgctcgcggc tcttaccagc
     5881 ctaacttcga tcactggacc gctgatcgtc acggcgattt atgccgcctc ggcgagcaca
     5941 tggaacgggt tggcatggat tgtaggcgcc gccctatacc ttgtctgcct ccccgcgttg
     6001 cgtcgcggtg catggagccg ggccacctcg acctgagaca attgtccaga taaaaaaaat
     6061 ccttagcttt cgctaaggat gatttctaaa atttttctgt catctcttcg ttattaatgt
     6121 ttgtaattga ctgaatatca acgcttattt aaatcagact gaagacttta tctctctgtc
     6181 ataaaactgt catattcctt acatataact gtcacctgtt tgtcctattt tgcttctcgt
     6241 agccaacaaa caatgcttta tgaatcctcc caggagacat tatgaaagtt atgcgtacca
     6301 ccgtcgcaac tgttgtcgcc gcgaccttat cgatgagtgc tttctctgtg tttgcagaag
     6361 caagcctgac aggtgcaggt gcaaccttcc ctgcgccggt gtatgccaaa tgggctgaca
     6421 cttaccagaa agaaaccggt aataaagtta actaccaggg tatcggttct tccggtggcg
     6481 taaaacagat tatcgctaat accgttgatt ttggtgcctc tgacgcgcc
//
