LOCUS       sequence-212090-        7803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eYFP Fluorescent tagging of bacteria at their Tn7-site.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7803)
  AUTHORS   Klausen M, Heydorn A, Ragas P, Lambertsen L, Aaes-Jorgensen A, Molin
            S, Tolker-Nielsen T
  TITLE     Biofilm formation by Pseudomonas aeruginosa wild type, flagella and 
            type IV pili mutants.
  JOURNAL   Mol Microbiol. 2003 Jun;48(6):1511-24.
  PUBMED    12791135
REFERENCE   2  (bases 1 to 7803)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Mol 
            Microbiol."; date: "2003-06"; volume: "48(6)"; pages: "1511-24"
COMMENT     Sequence Label: miniTn7(Gm)PA1/04/03-eyfp-a
FEATURES             Location/Qualifiers
     source          1..7803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    429..445
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     protein_bind    462..478
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     RBS             497..508
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(515..517,518..520,521..1234)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(560..581)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(821..840)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1168..1189
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      1257..1351
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             1395..2054
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1461..1480)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      2119..2205
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     oriT            3159..3268
                     /note="incP origin of transfer"
     CDS             3301..3672
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     complement(4340..4357)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4511..5099)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4591..4610)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5270..6061,6062..6130))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     5893..5912
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6131..6235)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6359..6381)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     6481..6500
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     6694..6716
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     6708..6725
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6709..6725
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  6770..6935
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        join(7059..7064,7065..7081,7082..7087)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             join(7276..7803,1..6)
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
ORIGIN
        1 acctaacaat tcgttcaagc cgagatcggc ttcccggccg cggagttgtt cggtaaattg
       61 tcacaacgcc gcggccaatt ctaattgggg accctagagg tccccttttt tattttaaaa
      121 attttttcac aaaacggttt acaagcataa agcttgctca atcaatcacc ggatcccgac
      181 gggcccggta ccgcggccgc gcgaattcga gctcggtacc atttatcagg gttattgtct
      241 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
      301 atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta
      361 taaaaatagg cgtatcacga ggccctttcg tcttcacctc gagaaaattt atcaaaaaga
      421 gtgttgactt gtgagcggat aacaatgata cttagattca attgtgagcg gataacaatt
      481 tcacacatct agaattaaag aggagaaatt aagcatgctg agcaagggcg aggagctgtt
      541 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
      601 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
      661 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccttcg gctacggcct
      721 gcagtgcttc gcccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
      781 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
      841 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
      901 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
      961 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     1021 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     1081 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agctaccagt ccgccctgag
     1141 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     1201 gatcactctc ggcatggacg agctgtacaa gtaataagct taattagctg agcttggact
     1261 cctgttgata gatccagtaa tgacctcaga actccatctg gatttgttca gaacgctcgg
     1321 ttgccgccgg gcgtttttta ttggtgagaa tccaagctag cttggcgaga ttttcaggag
     1381 ctaaggaagc taaaatggag aaaaaaatca ctggatatac caccgttgat atatcccaat
     1441 ggcatcgtaa agaacatttt gaggcatttc agtcagttgc tcaatgtacc tataaccaga
     1501 ccgttcagct ggatattacg gcctttttaa agaccgtaaa gaaaaataag cacaagtttt
     1561 atccggcctt tattcacatt cttgcccgcc tgatgaatgc tcatccggaa tttcgtatgg
     1621 caatgaaaga cggtgagctg gtgatatggg atagtgttca cccttgttac accgttttcc
     1681 atgagcaaac tgaaacgttt tcatcgctct ggagtgaata ccacgacgat ttccggcagt
     1741 ttctacacat atattcgcaa gatgtggcgt gttacggtga aaacctggcc tatttcccta
     1801 aagggtttat tgagaatatg tttttcgtct cagccaatcc ctgggtgagt ttcaccagtt
     1861 ttgatttaaa cgtggccaat atggacaact tcttcgcccc cgttttcacc atgggcaaat
     1921 attatacgca aggcgacaag gtgctgatgc cgctggcgat tcaggttcat catgccgttt
     1981 gtgatggctt ccatgtcggc agaatgctta atgaattaca acagtactgc gatgagtggc
     2041 agggcggggc gtaatttttt taaggcagtt attggtgccc ttaaacgcct ggggtaatga
     2101 ctctctagct tgaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt
     2161 tttatctgtt gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gctaggagct
     2221 tgcggccgcc ttaagacgcg tggatccccg ggaatttcga ccccacgccc ctctttaata
     2281 cgacgggcaa tttgcacttc agaaaatgaa gagtttgctt tagccataac aaaagtccag
     2341 tatgcttttt cacagcataa ctggactgat ttcagtttac aactattctg tctagtttaa
     2401 gactttattg tcatagttta gatctatttt gttcagttta agactttatt gtccgcccac
     2461 acccgcttac gcagggcatc catttattac tcaaccgtaa ccgattttgc caggttacgc
     2521 ggctggtcct ctagaggatc ctttttgtcc ggtgttgggt tgaaggtgaa gccggtcggg
     2581 gccgcagcgg gggccggctt ttcagccttg cccccctgct tcggccgccg tggctccggc
     2641 gtcttgggtg ccggcgcggg ttccgcagcc ttggcctgcg gtgcgggcac atcggcgggc
     2701 ttggccttga tgtgccgcct ggcgtgcgag cggaacgtct cgtaggagaa cttgaccttc
     2761 cccgtttccc gcatgtgctc ccaaatggtg acgagcgcat agccggacgc taacgccgcc
     2821 tcgacatccg ccctcaccgc caggaacgca accgcagcct catcacgccg gcgcttcttg
     2881 gccgcgcggg attcaaccca ctcggccagc tcgtcggtgt agctctttgg catcgtctct
     2941 cgcctgtccc ctcagttcag taatttcctg catttgcctg tttccagtcg gtagatattc
     3001 cacaaaacag cagggaagca gcgcttttcc gctgcataac cctgcttcgg ggtcattata
     3061 gcgatttttt cggtatatcc atcctttttc gcacgatata caggattttg ccaaagggtt
     3121 cgtgtagact ttccttggtg tatccaacgg cgtcagccgg gcaggatagg tgaagtaggc
     3181 ccacccgcga gcgggtgttc cttcttcact gtcccttatt cgcacctggc ggtgctcaac
     3241 gggaatcctg ctctgcgagg ctggccggct accgccggcg taacagatga gggcaagcgg
     3301 atggctgatg aaaccaagcc aaccaggaag ggcagcccac ctatcaaggt gtactgcctt
     3361 ccagacgaac gaagagcgat tgaggaaaag gcggcggcgg ccggcatgag cctgtcggcc
     3421 tacctgctgg ccgtcggcca gggctacaaa atcacgggcg tcgtggacta tgagcacgtc
     3481 cgcgagctgg cccgcatcaa tggcgacctg ggccgcctgg gcggcctgct gaaactctgg
     3541 ctcaccgacg acccgcgcac ggcgcggttc ggtgatgcca cgatcctcgc cctgctggcg
     3601 aagatcgaag agaagcagga cgagcttggc aaggtcatga tgggcgtggt ccgcccgagg
     3661 gcagagccat gactttttta gccgctaaaa cggccggggg gtgcgcgtga ttgccaagca
     3721 cgtccccatg cgctccatca agaagagcga cttcgcggag ctggtgaagt acatcaccga
     3781 cgagcaaggc aagaccgagc gcctgggtca cgtgcgcgtc acgaactgcg aggcaaacac
     3841 cctgcccgct gtcatggccg aggtgatggc gacccagcac ggcaacaccc gttccgaggc
     3901 cgacaagacc tatcacctgc tggttagctt ccgcgcggga gagaagcccg acgcggagac
     3961 gttgcgcgcg attgaggacc gcatctgcgc tgggcttggc ttcgccgagc atcagcgcgt
     4021 cagtgccgtg catcacgaca ccgacaacct gcacatccat atcgccatca acaagattca
     4081 cccgacccga aacaccatcc atgagccgta tcgggcctac cgcgccctcg ctgacctctg
     4141 cgcgacgctc gaacgggact acgggcttga gcgtgacaat cacgaaacgc ggcagcgcgt
     4201 ttccgagaac cgcgcgaacg acatggagcg gcacgcgggc gtggaaagcc tggtcggctg
     4261 gatcctctag agtcctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     4321 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4381 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4441 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4501 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4561 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4621 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4681 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4741 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4801 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4861 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4921 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4981 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     5041 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     5101 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     5161 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     5221 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     5281 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5341 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     5401 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     5461 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5521 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5581 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5641 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5701 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5761 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5821 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5881 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5941 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     6001 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     6061 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     6121 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     6181 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     6241 ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca ttaacctata
     6301 aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc
     6361 tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca
     6421 gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg
     6481 cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat
     6541 gcgtaaggag aaaataccgc atcaggcgcc attcgccatt caggctgcgc aactgttggg
     6601 aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg
     6661 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
     6721 ccagtgaatt cgagctcgcc caaacataac aggaagaaaa atgccccgct gtgggcggac
     6781 aaaatagttg ggaactggga ggggtggaaa tggagttttt aaggattatt tagggaagag
     6841 tgacaaaata gatgggaact gggtgtagcg tcgtaagcta atacgaaaat taaaaatgac
     6901 aaaatagttt ggaactagat ttcacttatc tggttggcct gcaggtcgac tagagtcggg
     6961 gatccggtga ttgattgagc aagctttatg cttgtaaacc gttttgtgaa aaaattttta
     7021 aaataaaaaa ggggacctct agggtcccca attacgaatt gacataagcc tgttcggttc
     7081 gtaaactgta atgcaagtag cgtatgcgct cacgcaactg gtccagaacc ttgaccgaac
     7141 gcagcggtgg taacggcgca gtggcggttt tcatggcttg ttatgactgt ttttttgtac
     7201 agtctatgcc tcgggcatcc aagcagcaag cgcgttacgc cgtgggtcga tgtttgatgt
     7261 tatggagcag caacgatgtt acgcagcagc aacgatgtta cgcagcaggg cagtcgccct
     7321 aaaacaaagt taggtggctc aagtatgggc atcattcgca catgtaggct cggccctgac
     7381 caagtcaaat ccatgcgggc tgctcttgat cttttcggtc gtgagttcgg agacgtagcc
     7441 acctactccc aacatcagcc ggactccgat tacctcggga acttgctccg tagtaagaca
     7501 ttcatcgcgc ttgctgcctt cgaccaagaa gcggttgttg gcgctctcgc ggcttacgtt
     7561 ctgcccaagt ttgagcagcc gcgtagtgag atctatatct atgatctcgc agtctccggc
     7621 gagcaccgga ggcagggcat tgccaccgcg ctcatcaatc tcctcaagca tgaggccaac
     7681 gcgcttggtg cttatgtgat ctacgtgcaa gcagattacg gtgacgatcc cgcagtggct
     7741 ctctatacaa agttgggcat acgggaagaa gtgatgcact ttgatatcga cccaagtacc
     7801 gcc
//
