LOCUS       sequence-464931-        6290 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  CCP expression in diverse gram-negative bacteria, with pre-cloned 
            Golden Gate Compatible (BsaI) restriction sites to facilitate 
            promoter swapping. .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 3697 to 3731)
  AUTHORS   Armstrong ZR, Alonso J, Stanton V, Patel N, Zogaj X, Cocioba SS, 
            Klose KE
  TITLE     Mobilizable shuttle vectors with fluorescent markers functional 
            across different species of bacteria.
  JOURNAL   Appl Environ Microbiol. 2025 May 12:e0004525. doi: 
            10.1128/aem.00045-25.
  PUBMED    40353662
REFERENCE   2  (bases 1 to 6290)
  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: "Appl 
            Environ Microbiol. 2025 May 12:e0004525. doi: 10.1128/aem.00045-25."
COMMENT     Sequence Label: pKEK3193
FEATURES             Location/Qualifiers
     source          1..6290
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     oriT            924..1033
                     /note="incP origin of transfer"
     CDS             1066..1437
                     /codon_start=1
                     /gene="traJ"
                     /product="oriT-recognizing protein"
                     /label=traJ
                     /translation="MADETKPTRKGSPPIKVYCLPDERRAIEEKAAAAGMSLSAYLLAV
                     GQGYKITGVVDYEHVRELARINGDLGRLGGLLKLWLTDDPRTARFGDATILALLAKIEE
                     KQDELGKVMMGVVRPRAEP"
     CDS             complement(2042..2719)
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGRKRR
                     ADPA"
     primer_bind     2634..2653
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     primer_bind     2911..2927
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2911..2927
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     RBS             3673..3684
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3697..3731)
                     /label=﻿BBa_J23100
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 1.0
                     "
                     /citation=[1]
     primer_bind     complement(3738..3754)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3808..4353
                     /direction=RIGHT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     4254..4273
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4470..4487
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 aaaaaaaaat ctgacaatat ataaagagaa tatataaaga gaatatctta ggggatttta
       61 aaaaaatccc acagactcaa agactttttt gactttttaa atcctagaaa ctatacttta
      121 agtacttatt taagtacatg gatttagatt atgcaaaccg ttaattattc aacttttaga
      181 aatgaactat ctgattcaat ggatagagta acaaaaaatc atagtcctat gattgtaact
      241 agaggttcaa aaaaagaagc agttgttatg atgtcgttag aggattttaa agctttgtta
      301 gcagcctttt tgtccggtgt tgggttgaag gtgaagccgg tcggggccgc agcgggggcc
      361 ggcttttcag ccttgccccc ctgcttcggc cgccgtggct ccggcgtctt gggtgccggc
      421 gcgggttccg cagccttggc ctgcggtgcg ggcacatcgg cgggcttggc cttgatgtgc
      481 cgcctggcgt gcgagcggaa cgtctcgtag gagaacttga ccttccccgt ttcccgcatg
      541 tgctcccaaa tggtgacgag cgcatagccg gacgctaacg ccgcctcgac atccgccctc
      601 accgccagga acgcaaccgc agcctcatca cgccggcgct tcttggccgc gcgggattca
      661 acccactcgg ccagctcgtc ggtgtagctc tttggcatcg tctctcgcct gtcccctcag
      721 ttcagtaatt tcctgcattt gcctgtttcc agtcggtaga tattccacaa aacagcaggg
      781 aagcagcgct tttccgctgc ataaccctgc ttcggggtca ttatagcgat tttttcggta
      841 tatccatcct ttttcgcacg atatacagga ttttgccaaa gggttcgtgt agactttcct
      901 tggtgtatcc aacggcgtca gccgggcagg ataggtgaag taggcccacc cgcgagcggg
      961 tgttccttct tcactgtccc ttattcgcac ctggcggtgc tcaacgggaa tcctgctctg
     1021 cgaggctggc cggctaccgc cggcgtaaca gatgagggca agcggatggc tgatgaaacc
     1081 aagccaacca ggaagggcag cccacctatc aaggtgtact gccttccaga cgaacgaaga
     1141 gcgattgagg aaaaggcggc ggcggccggc atgagcctgt cggcctacct gctggccgtc
     1201 ggccagggct acaaaatcac gggcgtcgtg gactatgagc acgtccgcga gctggcccgc
     1261 atcaatggcg acctgggccg cctgggcggc ctgctgaaac tctggctcac cgacgacccg
     1321 cgcacggcgc ggttcggtga tgccacgatc ctcgccctgc tggcgaagat cgaagagaag
     1381 caggacgagc ttggcaaggt catgatgggc gtggtccgcc cgagggcaga gccatgactt
     1441 ttttagccgc taaaacggcc ggggggtgcg cgtgattgcc aagcacgtcc ccatgcgctc
     1501 catcaagaag agcgacttcg cggagctggt gaagtacatc accgacgagc aaggcaagac
     1561 cgagcgcctg ggtcacgtgc gcgtcacgaa ctgcgaggca aacaccctgc ccgctgtcat
     1621 ggccgaggtg atggcgaccc agcacggcaa cacccgttcc gaggccgaca agacctatca
     1681 cctcctggtt agcttccgcg cgggagagaa gcccgacgcg gagacgttgc gcgcgattga
     1741 ggaccgcatc tgcgctgggc ttggcttcgc cgagcatcag cgcgtcagtg ccgtgcatca
     1801 cgacaccgac aacctgcaca tccatatcgc catcaacaag attcacccga cccgaaacac
     1861 catccatgag ccgtatcggg cctaccgcgc cctcgctgac ctctgcgcga cgctcgaacg
     1921 ggactacggg cttgagcgtg acaatcacga aacgcggcag cgcgtttccg agaaccgcgc
     1981 gaacgacatg gagcggcacg cgggcgtgga aagcctggtc ggctgctcgc gctttgttag
     2041 cagccggatc cgcgcggcgt ttacgccccc cctgccactc atcgcagtac tgttgtaatt
     2101 cattaagcat tctgccgaca tggaagccat cacaaacggc atgatgaacc tgaatcgcca
     2161 gcggcatcag caccttgtcg ccttgcgtat aatatttgcc catggtgaaa acgggggcga
     2221 agaagttgtc catattggcc acgtttaaat caaaactggt gaaactcacc cagggattgg
     2281 ctgagacgaa aaacatattc tcaataaacc ctttagggaa ataggccagg ttttcaccgt
     2341 aacacgccac atcttgcgaa tatatgtgta gaaactgccg gaaatcgtcg tggtattcac
     2401 tccagagcga tgaaaacgtt tcagtttgct catggaaaac ggtgtaacaa gggtgaacac
     2461 tatcccatat caccagctca ccgtctttca ttgccatacg gaattccgga tgagcattca
     2521 tcaggcgggc aagaatgtga ataaaggccg gataaaactt gtgcttattt ttctttacgg
     2581 tctttaaaaa ggccgtaata tccagctgaa cggtctggtt ataggtacat tgagcaactg
     2641 actgaaatgc ctcaaaatgt tctttacgat gccattggga tatatcaacg gtggtatatc
     2701 cagtgatttt tttctccata tgtttagtct cctttttttg ttataaatat tttatacaca
     2761 aaaatcttcg ttatttaaat aagttttata gctatttttt caagatacaa aactattaat
     2821 tagttataat taggttatta gcttaaaatt ataaaccaaa tacgatgagt gacaacccaa
     2881 aagatctcga tcctctacgc cgttggggag caggaaacag ctatgacggt accaaaaaaa
     2941 accccgccct gtcaggggcg gggttttttt ttgagctctt attatgggag agccttcggc
     3001 agcggtgaga aatggccttc ggctgcctcc tgctggtccc aagttgtgaa atctttatcg
     3061 tgtttggtca gttccaggcg gtgttcaacg aaatgaaatt ctggcagctt gacaggtttt
     3121 ttggctttat acgtcgtttt gaaatcaaca cgatgatgac cgcctccttt cagtttaaaa
     3181 accattgcag tgtcactttt taacatcccg tcacgcacgt acagtgtttc cacgctcggc
     3241 tcccatccag caatggtttt ttgcattacg ggcccatccg gaggcaggtt cacaccatga
     3301 aatttaacat catagatgaa acagttctct tgcaagttta tgctggagtg attactggcg
     3361 gtgcatacgc caccatcttc aaaaatcatc gaccgctccc aagacatacc ttcgggaaaa
     3421 gactgtttgt ggtagtctgg gatatccgct ggatacttat taaacggttt gcttccatac
     3481 atgtggctag gtgtaagaat atgaacggaa aacggcagag gcgcgccttt ggtgactgtc
     3541 attttcatgt gctgtaaccc ttcgtacggc ttgccttcac ctttaccctc gatcgtaaaa
     3601 tcatggccat tgacagtgcc ctcaaggtgc aacttggtct tcattacctg cttaatcact
     3661 gccatgggac tctttctcct ctttaatcga gaccaagcta gcactgtacc taggactgag
     3721 ctagccgtca agaattcact ggccgtcgtt ttacggtctc agtcgactac tctagaaata
     3781 ttttatctga ttaataagat gatcttcttg agatcgtttt ggtctgcgcg taatctcttg
     3841 ctctgaaaac gaaaaaaccg ccttgcaggg cggtttttcg aaggttctct gagctaccaa
     3901 ctctttgaac cgaggtaact ggcttggagg agcgcagtca ccaaaacttg tcctttcagt
     3961 ttagccttaa ccggcgcatg acttcaagac taactcctct aaatcaatta ccagtggctg
     4021 ctgccagtgg tgcttttgca tgtctttccg ggttggactc aagacgatag ttaccggata
     4081 aggcgcagcg gtcggactga acggggggtt cgtgcataca gtccagcttg gagcgaactg
     4141 cctacccgga actgagtgtc aggcgtggaa tgagacaaac gcggccataa cagcggaatg
     4201 acaccggtaa accgaaaggc aggaacagga gagcgcacga gggagccgcc agggggaaac
     4261 gcctggtatc tttatagtcc tgtcgggttt cgccaccact gatttgagcg tcagatttcg
     4321 tgatgcttgt caggggggcg gagcctatgg aaaaacggct ttgccgcggc cctctcactt
     4381 ccctgttaag tatcttcctg gcatcttcca ggaaatctcc gccccgttcg taagccattt
     4441 ccgctcgccg cagtcgaacg accgagcgta gcgagtcagt gagcgaggaa gcggaatata
     4501 tcctgtatca catattctgc tgacgcaccg gtgcagcctt ttttctcctg ccacatgaag
     4561 cacttcactg acaccctcat cagtgccaac atagtaagcc agtatacact ccgctagcgc
     4621 tgatgtccgg cggtgctttt gccgttacgc accaccccgt cagtagctga acaggaggga
     4681 cagctaaagc tagaaatgta ccaaatttta cctattttag gatacttttc atgaagctct
     4741 tctatttttc cccagtcctt taatagtcta cctttagagt ctcgtaaata gttatctttg
     4801 tgacaggggc ctcttttatc ttttttaatg taactatatg ttattccaac gtcactatta
     4861 ctattatcca aatctttttt agcatgccag taagaacttt cataacttaa ctctatcttt
     4921 cgacctcttt gatatacaac aataaagcta tagccagtag taacaacctg ttttactttt
     4981 gttaaatcta ttaacttctt atttattttt ttatgttttt ttgaaaattt aaatatttct
     5041 atattcattc ctacacttcc tcaaatccaa atggtagctt atgattctct tctggtttct
     5101 tttctaattt ttttatattt gcaataaaaa ctctttttct atctttgatt tttttattgt
     5161 cccaattcct ccaagtatca tcacaaaccc tttcaatatc atgtaaatga tgatgtctaa
     5221 atattgatct gacataatac agatctaggt ctagttcatc acttaacaca acttctctaa
     5281 gtctttcaga tgcttcgatt ggtatgtaat cctctttatt tttagtatct aaaggctttt
     5341 gcttaaattc ttcttctgtc tctgctacct tactaactgt aaacttgata tttgtaatct
     5401 tacgaccatg ttttctgtga tgatccttgt catcataggt tacaaaaata tccgataatt
     5461 gattaatctc ttctagtgct ggtaatagga acttattttt aaaatttgaa tatctgttgc
     5521 tgtaactttt aggtaaatca aaatcattaa tcatatcatc gacatacaat acgcaatcaa
     5581 ctatattagc ataccctgct tgttcgccta atttgctttt gagaagtaag tataatctgc
     5641 ttgaatactt acttttaaat gaaaatagta actgtctttc tgctttagta aagtactctt
     5701 gtagttgtat catgtgtggc attaatgacc aatgaaactc gcaaattaaa gcactgcttt
     5761 tagggtctgc ttcaatatat gcaaaccagt tagctatctt cgtttgttct ttattcagcc
     5821 atactggctt agacattatt gagtgcatta attgcttcaa tctcactctg ttatgcttaa
     5881 cccctgtagc tttttcaaga tcagataggc ttatcttata cctgtgaaac tctttatctt
     5941 ctcttttaac cattgaggca actaagaata ttaagttttg ttcttctttt gtaaggctat
     6001 actttcctgc aacaagagta ttagacatag ctatttcttt gccagcattt acatttttaa
     6061 cttctttcat agaactagag tcattatctc gatatacaaa ttctataaaa cttctattag
     6121 taaaacaact acttcataaa aaaaagtagt tttaacgata caaaaagtag ttttaaattc
     6181 aaaaagtgat acaaaaagta gttttaaatt caaaaagtga tacaaaaagt agttttaaat
     6241 tttttaaaaa agtgcttcaa agccttatgt agcaatactt acagaggatt
//
