LOCUS       sequence-329150-        7803 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  eCFP 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-ecfp-a
FEATURES             Location/Qualifiers
     source          1..7803
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     55..77
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     69..86
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     70..86
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     mobile_element  131..296
                     /mobile_element_type="transposon:Tn7"
                     /label=Tn7L
                     /note="mini-Tn7 element (left end of the Tn7 transposon)"
     promoter        join(420..425,426..442,443..448)
                     /gene="intI1 (promoter lies within the coding sequence)"
                     /label=Pc promoter
                     /note="class 1 integron promoter"
     CDS             637..1170
                     /codon_start=1
                     /gene="aacC1"
                     /product="gentamycin acetyltransferase"
                     /label=GmR
                     /note="confers resistance to gentamycin"
                     /translation="MLRSSNDVTQQGSRPKTKLGGSSMGIIRTCRLGPDQVKSMRAALD
                     LFGREFGDVATYSQHQPDSDYLGNLLRSKTFIALAAFDQEAVVGALAAYVLPKFEQPRS
                     EIYIYDLAVSGEHRRQGIATALINLLKHEANALGAYVIYVQADYGDDPAVALYTKLGIR
                     EEVMHFDIDPSTAT"
     protein_bind    1593..1609
                     /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    1626..1642
                     /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             1661..1672
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             join(1679..1681,1682..1684,1685..2398)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MLSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1724..1745)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1985..2004)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2332..2353
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     terminator      2421..2515
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             2559..3218
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(2625..2644)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     terminator      3283..3369
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     oriT            4323..4432
                     /note="incP origin of transfer"
     CDS             4465..4836
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     primer_bind     complement(5504..5521)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5675..6263)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5755..5774)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6434..7225,7226..7294))
                     /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     7057..7076
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7295..7399)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7523..7545)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7645..7664
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 tggcgaaagg gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt
       61 cacgacgttg taaaacgacg gccagtgaat tcgagctcgc ccaaacataa caggaagaaa
      121 aatgccccgc tgtgggcgga caaaatagtt gggaactggg aggggtggaa atggagtttt
      181 taaggattat ttagggaaga gtgacaaaat agatgggaac tgggtgtagc gtcgtaagct
      241 aatacgaaaa ttaaaaatga caaaatagtt tggaactaga tttcacttat ctggttggcc
      301 tgcaggtcga ctagagtcgg ggatccggtg attgattgag caagctttat gcttgtaaac
      361 cgttttgtga aaaaattttt aaaataaaaa aggggacctc tagggtcccc aattacgaat
      421 tgacataagc ctgttcggtt cgtaaactgt aatgcaagta gcgtatgcgc tcacgcaact
      481 ggtccagaac cttgaccgaa cgcagcggtg gtaacggcgc agtggcggtt ttcatggctt
      541 gttatgactg tttttttgta cagtctatgc ctcgggcatc caagcagcaa gcgcgttacg
      601 ccgtgggtcg atgtttgatg ttatggagca gcaacgatgt tacgcagcag caacgatgtt
      661 acgcagcagg gcagtcgccc taaaacaaag ttaggtggct caagtatggg catcattcgc
      721 acatgtaggc tcggccctga ccaagtcaaa tccatgcggg ctgctcttga tcttttcggt
      781 cgtgagttcg gagacgtagc cacctactcc caacatcagc cggactccga ttacctcggg
      841 aacttgctcc gtagtaagac attcatcgcg cttgctgcct tcgaccaaga agcggttgtt
      901 ggcgctctcg cggcttacgt tctgcccaag tttgagcagc cgcgtagtga gatctatatc
      961 tatgatctcg cagtctccgg cgagcaccgg aggcagggca ttgccaccgc gctcatcaat
     1021 ctcctcaagc atgaggccaa cgcgcttggt gcttatgtga tctacgtgca agcagattac
     1081 ggtgacgatc ccgcagtggc tctctataca aagttgggca tacgggaaga agtgatgcac
     1141 tttgatatcg acccaagtac cgccacctaa caattcgttc aagccgagat cggcttcccg
     1201 gccgcggagt tgttcggtaa attgtcacaa cgccgcggcc aattctaatt ggggacccta
     1261 gaggtcccct tttttatttt aaaaattttt tcacaaaacg gtttacaagc ataaagcttg
     1321 ctcaatcaat caccggatcc cgacgggccc ggtaccgcgg ccgcgcgaat tcgagctcgg
     1381 taccatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     1441 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac
     1501 cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtcttca
     1561 cctcgagaaa atttatcaaa aagagtgttg acttgtgagc ggataacaat gatacttaga
     1621 ttcaattgtg agcggataac aatttcacac atctagaatt aaagaggaga aattaagcat
     1681 gctgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg
     1741 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg
     1801 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct
     1861 cgtgaccacc ctgacctggg gcgtgcagtg cttcagccgc taccccgacc acatgaagca
     1921 gcacgacttc ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt
     1981 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt
     2041 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa
     2101 gctggagtac aactacatca gccacaacgt ctatatcacc gccgacaagc agaagaacgg
     2161 catcaaggcc aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga
     2221 ccactaccag cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta
     2281 cctgagcacc cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct
     2341 gctggagttc gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagtaata
     2401 agcttaatta gctgagcttg gactcctgtt gatagatcca gtaatgacct cagaactcca
     2461 tctggatttg ttcagaacgc tcggttgccg ccgggcgttt tttattggtg agaatccaag
     2521 ctagcttggc gagattttca ggagctaagg aagctaaaat ggagaaaaaa atcactggat
     2581 ataccaccgt tgatatatcc caatggcatc gtaaagaaca ttttgaggca tttcagtcag
     2641 ttgctcaatg tacctataac cagaccgttc agctggatat tacggccttt ttaaagaccg
     2701 taaagaaaaa taagcacaag ttttatccgg cctttattca cattcttgcc cgcctgatga
     2761 atgctcatcc ggaatttcgt atggcaatga aagacggtga gctggtgata tgggatagtg
     2821 ttcacccttg ttacaccgtt ttccatgagc aaactgaaac gttttcatcg ctctggagtg
     2881 aataccacga cgatttccgg cagtttctac acatatattc gcaagatgtg gcgtgttacg
     2941 gtgaaaacct ggcctatttc cctaaagggt ttattgagaa tatgtttttc gtctcagcca
     3001 atccctgggt gagtttcacc agttttgatt taaacgtggc caatatggac aacttcttcg
     3061 cccccgtttt caccatgggc aaatattata cgcaaggcga caaggtgctg atgccgctgg
     3121 cgattcaggt tcatcatgcc gtttgtgatg gcttccatgt cggcagaatg cttaatgaat
     3181 tacaacagta ctgcgatgag tggcagggcg gggcgtaatt tttttaaggc agttattggt
     3241 gcccttaaac gcctggggta atgactctct agcttgaggc atcaaataaa acgaaaggct
     3301 cagtcgaaag actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
     3361 aggacaaatc cgccgctagg agcttgcggc cgccttaaga cgcgtggatc cccgggaatt
     3421 tcgaccccac gcccctcttt aatacgacgg gcaatttgca cttcagaaaa tgaagagttt
     3481 gctttagcca taacaaaagt ccagtatgct ttttcacagc ataactggac tgatttcagt
     3541 ttacaactat tctgtctagt ttaagacttt attgtcatag tttagatcta ttttgttcag
     3601 tttaagactt tattgtccgc ccacacccgc ttacgcaggg catccattta ttactcaacc
     3661 gtaaccgatt ttgccaggtt acgcggctgg tcctctagag gatccttttt gtccggtgtt
     3721 gggttgaagg tgaagccggt cggggccgca gcgggggccg gcttttcagc cttgcccccc
     3781 tgcttcggcc gccgtggctc cggcgtcttg ggtgccggcg cgggttccgc agccttggcc
     3841 tgcggtgcgg gcacatcggc gggcttggcc ttgatgtgcc gcctggcgtg cgagcggaac
     3901 gtctcgtagg agaacttgac cttccccgtt tcccgcatgt gctcccaaat ggtgacgagc
     3961 gcatagccgg acgctaacgc cgcctcgaca tccgccctca ccgccaggaa cgcaaccgca
     4021 gcctcatcac gccggcgctt cttggccgcg cgggattcaa cccactcggc cagctcgtcg
     4081 gtgtagctct ttggcatcgt ctctcgcctg tcccctcagt tcagtaattt cctgcatttg
     4141 cctgtttcca gtcggtagat attccacaaa acagcaggga agcagcgctt ttccgctgca
     4201 taaccctgct tcggggtcat tatagcgatt ttttcggtat atccatcctt tttcgcacga
     4261 tatacaggat tttgccaaag ggttcgtgta gactttcctt ggtgtatcca acggcgtcag
     4321 ccgggcagga taggtgaagt aggcccaccc gcgagcgggt gttccttctt cactgtccct
     4381 tattcgcacc tggcggtgct caacgggaat cctgctctgc gaggctggcc ggctaccgcc
     4441 ggcgtaacag atgagggcaa gcggatggct gatgaaacca agccaaccag gaagggcagc
     4501 ccacctatca aggtgtactg ccttccagac gaacgaagag cgattgagga aaaggcggcg
     4561 gcggccggca tgagcctgtc ggcctacctg ctggccgtcg gccagggcta caaaatcacg
     4621 ggcgtcgtgg actatgagca cgtccgcgag ctggcccgca tcaatggcga cctgggccgc
     4681 ctgggcggcc tgctgaaact ctggctcacc gacgacccgc gcacggcgcg gttcggtgat
     4741 gccacgatcc tcgccctgct ggcgaagatc gaagagaagc aggacgagct tggcaaggtc
     4801 atgatgggcg tggtccgccc gagggcagag ccatgacttt tttagccgct aaaacggccg
     4861 gggggtgcgc gtgattgcca agcacgtccc catgcgctcc atcaagaaga gcgacttcgc
     4921 ggagctggtg aagtacatca ccgacgagca aggcaagacc gagcgcctgg gtcacgtgcg
     4981 cgtcacgaac tgcgaggcaa acaccctgcc cgctgtcatg gccgaggtga tggcgaccca
     5041 gcacggcaac acccgttccg aggccgacaa gacctatcac ctgctggtta gcttccgcgc
     5101 gggagagaag cccgacgcgg agacgttgcg cgcgattgag gaccgcatct gcgctgggct
     5161 tggcttcgcc gagcatcagc gcgtcagtgc cgtgcatcac gacaccgaca acctgcacat
     5221 ccatatcgcc atcaacaaga ttcacccgac ccgaaacacc atccatgagc cgtatcgggc
     5281 ctaccgcgcc ctcgctgacc tctgcgcgac gctcgaacgg gactacgggc ttgagcgtga
     5341 caatcacgaa acgcggcagc gcgtttccga gaaccgcgcg aacgacatgg agcggcacgc
     5401 gggcgtggaa agcctggtcg gctggatcct ctagagtcct gcattaatga atcggccaac
     5461 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5521 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5581 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5641 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5701 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5761 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5821 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5881 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5941 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     6001 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     6061 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     6121 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     6181 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     6241 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6301 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6361 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6421 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6481 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6541 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6601 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6661 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6721 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6781 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6841 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6901 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6961 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     7021 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     7081 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     7141 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     7201 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     7261 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7321 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7381 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     7441 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtctcgcgc
     7501 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     7561 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     7621 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     7681 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     7741 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     7801 agc
//
