LOCUS       SEGA055                5534 bp ds-DNA     linear   UNA 17-JUN-2021
DEFINITION  LMBP 12891.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 5534)
  AUTHORS   Morten Norholm Group
  TITLE     Direct Submission
  JOURNAL   Exported Jun 17, 2021 from SnapGene 4.2.11
COMMENT     SEGA landing pad: PRha-high-YebF-sfGFP-T-tetA-S2P11-S3P22-rrnC
FEATURES             Location/Qualifiers
     source          1..5534
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     source          2656..2718
                     /organism="Escherichia coli str. K-12 substr. MG1655"
                     /strain="K-12"
                     /sub_strain="MG1655"
                     /mol_type="genomic DNA"
                     /db_xref="taxon:511145"
     CDS             1..528
                     /codon_start=3
                     /label=glmS
                     /translation="VHGLQALPSRIEQMLSQDKRIEALAEDFSDKHHALFLGRGDQYPI
                     ALEGALKLKEISYIHAEAYAAGELKHGPLALIDADMPVIVVAPNNELLEKLKSNIEEVR
                     ARGGQLYVFADQDAGFVSSDNMHIIEMPHVEEVIAPIFYTVPLQLLAYHVALIKGTDVD
                     QPRNLAKSVTVE"
     misc_feature    538..594
                     /label=L3S2P11 Terminator
     CDS             complement(607..1452)
                     /codon_start=1
                     /gene="E. coli rhaR"
                     /product="transcriptional activator of rhaR and rhaS"
                     /label=rhaR
                     /note="needed for rhamnose-dependent activation of the rhaB
                     promoter"
                     /translation="MAHQLKLLKDDFFASDQQAVAVADRYPQDVFAEHTHDFCELVIVW
                     RGNGLHVLNDRPYRITRGDLFYIHADDKHSYASVNDLVLQNIIYCPERLKLNLDWQGAI
                     PGFNASAGQPHWRLGSMGMAQARQVIGQLEHESSQHVPFANEMAELLFGQLVMLLNRHR
                     YTSDSLPPTSSETLLDKLITRLAASLKSPFALDKFCDEASCSERVLRQQFRQQTGMTIN
                     QYLRQVRVCHAQYLLQHSRLLISDISTECGFEDSNYFSVVFTRETGMTPSQWRHLNSQK
                     D"
     CDS             complement(1526..2362)
                     /codon_start=1
                     /gene="E. coli rhaS"
                     /product="positive regulator of the rhaB promoter"
                     /label=rhaS
                     /note="activated by L-rhamnose"
                     /translation="MTVLHSVDFFPSGNASVAIEPRLPQADFPEHHHDFHEIVIVEHGT
                     GIHVFNGQPYTITGGTVCFVRDHDRHLYEHTDNLCLTNVLYRSPDRFQFLAGLNQLLPQ
                     ELDGQYPSHWRVNHSVLQQVRQLVAQMEQQEGENDLPSTASREILFMQLLLLLRKSSLQ
                     ENLENSASRLNLLLAWLEDHFADEVNWDAVADQFSLSLRTLHRQLKQQTGLTPQRYLNR
                     LRLMKARHLLRHSEASVTDIAYRCGFSDSNHFSTLFRREFNWSPRDIRQGRDGFLQ"
     promoter        2508..2626
                     /label=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             2638..2649
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     misc_feature    2650..2664
                     /label=TIR
     CDS             2656..2718
                     /codon_start=1
                     /transl_table=11
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /product="extracellular Colicin M immunity family protein"
                     /label=yebF
                     /db_xref="GI:145698278"
                     /db_xref="ASAP:ABE-0006141"
                     /db_xref="UniProtKB/Swiss-Prot:P33219"
                     /db_xref="EcoGene:EG11807"
                     /db_xref="GeneID:946363"
                     /protein_id="NP_416361.2"
                     /translation="MKKRGAFLGLLLVSACASVFA"
     gene            2656..2718
                     /gene="yebF"
                     /gene_synonym="ECK1848; JW1836"
                     /locus_tag="b1847"
                     /label=yebF
                     /db_xref="EcoGene:EG11807"
                     /db_xref="GeneID:946363"
     CDS             2719..3009
                     /codon_start=1
                     /label=YebF
                     /translation="ANNETSKSVTFPKCEDLDAAGIAASVKRDYQQNRVARWADDQKIV
                     GQADPVAWVSLQDIQGKDDKWSVPLTVRGKSADIHYQVSVDCKAGMAEYQRR"
     CDS             3010..3720
                     /codon_start=1
                     /label=sfGFP
                     /translation="RKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTLKF
                     ICTTGKLPVPWPTLVTTLTYGVQCFARYPDHMKQHDFFKSAMPEGYVQERTISFKDDGT
                     YKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKAN
                     FKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSVLSKDPNEKRDHMVLLEF
                     VTAAGITHGMDELYK"
     misc_feature    3724..3772
                     /label=L3S3P22 Terminator (synthetic)
     promoter        3783..3811
                     /label=Promoter 3BCD2
     CDS             3851..5041
                     /codon_start=1
                     /gene="tet"
                     /product="tetracycline efflux protein"
                     /label=TcR
                     /note="confers resistance to tetracycline"
                     /translation="MKSNNALIVILGTVTLDAVGIGLVMPVLPGLLRDIVHSDSIASHY
                     GVLLALYALMQFLCAPVLGALSDRFGRRPVLLASLLGATIDYAIMATTPVLWILYAGRI
                     VAGITGATGAVAGAYIADITDGEDRARHFGLMSACFGVGMVAGPVAGGLLGAISLHAPF
                     LAAAVLNGLNLLLGCFLMQESHKGERRPMPLRAFNPVSSFRWARGMTIVAALMTVFFIM
                     QLVGQVPAALWVIFGEDRFRWSATMSGLSLAVFGILHALAQAFVTGPATKRFGEKQAII
                     AGMAADALGYVLLAFATRGWMAFPIMILLASGGIGMPALQAMLSRQVDDDHQGQLQGSL
                     AALTSLTSITGPLIVTAIYAASASTWNGLAWIVGAALYLVCLPALRRGAWSRATST"
     misc_feature    5022..5071
                     /label=HR3
     terminator      5052..5092
                     /label=rrnC
     misc_feature    5093..5142
                     /label=homology
     CDS             5287..5534
                     /codon_start=1
                     /label=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 gtccgcgttg
      601 cgcttaatct ttctgcgaat tgagatgacg ccactggctg ggcgtcatcc cggtttcccg
      661 ggtaaacacc accgaaaaat agttactatc ttcaaagcca cattcggtcg aaatatcact
      721 gattaacagg cggctatgct ggagaagata ttgcgcatga cacactctga cctgtcgcag
      781 atattgattg atggtcattc cagtctgctg gcgaaattgc tgacgcaaaa cgcgctcact
      841 gcacgatgcc tcatcacaaa atttatccag cgcaaaggga cttttcaggc tagccgccag
      901 ccgggtaatc agcttatcca gcaacgtttc gctggatgtt ggcggcaacg aatcactggt
      961 gtaacgatgg cgattcagca acatcaccaa ctgcccgaac agcaactcag ccatttcgtt
     1021 agcaaacggc acatgctgac tactttcatg ctcaagctga ccaataacct gccgcgcctg
     1081 cgccatcccc atgctaccta agcgccagtg tggttgccct gcgctggcgt taaatcccgg
     1141 aatcgccccc tgccagtcaa gattcagctt cagacgctcc gggcaataaa taatattctg
     1201 caaaaccaga tcgttaacgg aagcgtagga gtgtttatca tcagcatgaa tgtaaaagag
     1261 atcgccacgg gtaatgcgat aagggcgatc gttgagtaca tgcaggccat taccgcgcca
     1321 gacaatcacc agctcacaaa aatcatgtgt atgttcagca aagacatctt gcggataacg
     1381 gtcagccaca gcgactgcct gctggtcgct ggcaaaaaaa tcatctttga gaagttttaa
     1441 ctgatgcgcc accgtggcta cctcggccag agaacgaagt tgattattcg caatatggcg
     1501 tacaaatacg ttgagaagat tcgcgttatt gcagaaagcc atcccgtccc tggcgaatat
     1561 cacgcggtga ccagttaaac tctcggcgaa aaagcgtcga aaagtggtta ctgtcgctga
     1621 atccacagcg ataggcgatg tcagtaacgc tggcctcgct gtggcgtagc agatgtcggg
     1681 ctttcatcag tcgcaggcgg ttcaggtatc gctgaggcgt cagtcccgtt tgctgcttaa
     1741 gctgccgatg tagcgtacgc agtgaaagag aaaattgatc cgccacggca tcccaattca
     1801 cctcatcggc aaaatggtcc tccagccagg ccagaagcaa gttgagacgt gatgcgctgt
     1861 tttccaggtt ctcctgcaaa ctgcttttac gcagcaagag cagtaattgc ataaacaaga
     1921 tctcgcgact ggcggtcgag ggtaaatcat tttccccttc ctgctgttcc atctgtgcaa
     1981 ccagctgtcg cacctgctgc aatacgctgt ggttaacgcg ccagtgagac ggatactgcc
     2041 catccagctc ttgtggcagc aactgattca gcccggcgag aaactgaaat cgatccggcg
     2101 agcgatacag cacattggtc agacacagat tatcggtatg ttcatacaga tgccgatcat
     2161 gatcgcgtac gaaacagacc gtgccaccgg tgatggtata gggctgccca ttaaacacat
     2221 gaatacccgt gccatgttcg acaatcacaa tttcatgaaa atcatgatga tgttcaggaa
     2281 aatccgcctg cgggagccgg ggttctatcg ccacggacgc gttaccagac ggaaaaaaat
     2341 ccacactatg taatacggtc atactggcct cctgatgtcg tcaacacggc gaaatagtaa
     2401 tcacgaggtc aggttcttac cttaaatttt cgacggaaaa ccacgtaaaa aacgtcgatt
     2461 tttcaagata cagcgtgaat tttcaggaaa tgcggtgagc atcacatcac cacaattcag
     2521 caaattgtga acatcatcac gttcatcttt ccctggttgc caatggccca ttttcttgtc
     2581 agtaacgaga aggtcgcgaa tccaggcgct ttttagactg gtcgtatact agagattaaa
     2641 gaggagaaag gggctatgaa aaagagaggg gcgtttttag ggctgttgtt ggtttctgcc
     2701 tgcgcatcag ttttcgctgc gaataacgaa acgagcaaga gcgtcacctt cccgaagtgc
     2761 gaggatctgg acgcagcggg cattgctgcc agcgtgaaac gtgactacca acagaaccgc
     2821 gtggctcgtt gggcagacga ccagaagatc gtcggtcaag cggacccggt tgcctgggtt
     2881 tccctgcaag atatccaggg caaagatgat aaatggagcg tcccgctgac cgttcgtggt
     2941 aagagcgcgg acattcacta tcaggtgagc gtcgattgca aagccggcat ggcggagtac
     3001 caacgccgcc gtaaaggcga agagctgttc actggtgtcg tccctattct ggtggaactg
     3061 gatggtgatg tcaacggtca taagttttcc gtgcgtggcg agggtgaagg tgacgcaact
     3121 aatggtaaac tgacgctgaa gttcatctgt actactggta aactgccggt accttggccg
     3181 actctggtaa cgacgctgac ttatggtgtt cagtgctttg ctcgttatcc ggaccacatg
     3241 aagcagcatg acttcttcaa gtccgccatg ccggaaggct atgtgcagga acgcacgatt
     3301 tcctttaagg atgacggcac gtacaaaacg cgtgcggaag tgaaatttga aggcgatacc
     3361 ctggtaaacc gcattgagct gaaaggcatt gactttaaag aagacggcaa tatcctgggc
     3421 cataagctgg aatacaattt taacagccac aatgtttaca tcaccgccga taaacaaaaa
     3481 aatggcatta aagcgaattt taaaattcgc cacaacgtgg aggatggcag cgtgcagctg
     3541 gctgatcact accagcaaaa cactccaatc ggtgatggtc ctgttctgct gccagacaat
     3601 cactatctga gcacgcaaag cgttctgtct aaagatccga acgagaaacg cgatcacatg
     3661 gttctgctgg agttcgtaac cgcagcgggc atcacgcatg gtatggatga actgtacaaa
     3721 tgaccaatta ttgaaggccg ctaacgcggc ctttttttgt ttctggtctc ccaaaaattt
     3781 atttgcttat taatcatccg gctcgtataa tgtgtggagg gcccaagttc acttaaaaag
     3841 gagaatagtt atgaaatcga acaatgcgct catcgtcatc ctcggcaccg tcaccctgga
     3901 tgctgtaggc ataggcttgg ttatgccggt actgccgggc ctcttgcggg atatcgtcca
     3961 ttccgacagc atcgccagtc actatggcgt gctgctagcg ctatatgcgt tgatgcaatt
     4021 tctatgcgca cccgttctcg gagcactgtc cgaccgcttt ggccgccgcc cagtcctgct
     4081 cgcttcgcta cttggagcca ctatcgacta cgcgatcatg gcgaccacac ccgtcctgtg
     4141 gattctctac gccggacgca tcgtggccgg catcaccggc gccacaggtg cggttgctgg
     4201 cgcctatatc gccgacatca ccgatgggga agatcgggct cgccacttcg ggctcatgag
     4261 cgcttgtttc ggcgtgggta tggtggcagg ccccgtggcc gggggactgt tgggcgccat
     4321 ctccctgcac gcaccattcc ttgcggcggc ggtgctcaac ggcctcaacc tactactggg
     4381 ctgcttccta atgcaggagt cgcataaggg agagcgccgt ccgatgccct tgagagcctt
     4441 caacccagtc agctccttcc ggtgggcgcg gggcatgacc attgtggccg cacttatgac
     4501 tgtcttcttt atcatgcaac tcgtaggaca ggtgccggca gcgctctggg tcattttcgg
     4561 cgaggaccgc tttcgctgga gcgcgacgat gagcggcctg tcgcttgcgg tattcggaat
     4621 cttgcacgcc ctcgctcaag ccttcgtcac tggtcccgcc accaaacgtt tcggcgagaa
     4681 gcaggccatt atcgccggca tggcggccga cgcgctgggc tacgtcttgc tggcgttcgc
     4741 gacgcgaggc tggatggcct tccccattat gattcttctc gcttccggcg gcatcgggat
     4801 gcccgcgttg caggccatgc tgtccaggca ggtagatgac gaccatcagg gacagcttca
     4861 aggatcgctc gcggctctta ccagcctaac ttcgatcact ggaccgctga tcgtcacggc
     4921 gatttatgcc gcctcggcga gcacatggaa cgggttggca tggattgtag gcgccgccct
     4981 ataccttgtc tgcctccccg cgttgcgtcg cggtgcatgg agccgggcca cctcgacctg
     5041 agacaattgt ccagataaaa aaaatcctta gctttcgcta aggatgattt ctaaaatttt
     5101 tctgtcatct cttcgttatt aatgtttgta attgactgaa tatcaacgct tatttaaatc
     5161 agactgaaga ctttatctct ctgtcataaa actgtcatat tccttacata taactgtcac
     5221 ctgtttgtcc tattttgctt ctcgtagcca acaaacaatg ctttatgaat cctcccagga
     5281 gacattatga aagttatgcg taccaccgtc gcaactgttg tcgccgcgac cttatcgatg
     5341 agtgctttct ctgtgtttgc agaagcaagc ctgacaggtg caggtgcaac cttccctgcg
     5401 ccggtgtatg ccaaatgggc tgacacttac cagaaagaaa ccggtaataa agttaactac
     5461 cagggtatcg gttcttccgg tggcgtaaaa cagattatcg ctaataccgt tgattttggt
     5521 gcctctgacg cgcc
//
