LOCUS       sequence-464933-        6293 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 3700 to 3734)
  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 6293)
  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: pKEK3199
FEATURES             Location/Qualifiers
     source          1..6293
                     /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             3676..3687
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3700..3734)
                     /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(3741..3757)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3811..4356
                     /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     4257..4276
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4473..4490
                     /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 ttgagctctc attaagttgc tttctcaggc
     3001 acatcagagt aacgggccac agccgactca tactgttcat aaaatttccc ctgttccgcc
     3061 tttaagatct caatgcggtg gtcgccaaag tgaaactcag gcatgttgac cgcgttggag
     3121 ggcttacggc tacggtatgt tgtacgtaag tgcgacgtta agtgattacc atcggtacat
     3181 ttcagtgcca tcagactttg gccgcacaat accccgtcac gtgggtaaag catctcaaca
     3241 cacggctccc agcctgcagt tttcttttgc ataacgggtc cgttggcagg gaaattcgtc
     3301 ccaagcactt tcactttgta aatcaagcaa ttgccctcaa gacttgtatc ctgatgtgcg
     3361 gtcaatactc cgccatcctc gaagatctgt gtacgctccc aggtgaaccc ttcggggaac
     3421 gactctttaa aatagtctgg gatgccgctg acatgtttaa taaaggtttt gctcccatac
     3481 atgcaacatg gtgccagaat atcgaaagcg aacggtaaag gtccaccctc cgtgacgcga
     3541 atacgcatgt tttgagttcc ctcaaacggt ttcccctcac cttcccccac acacttaaag
     3601 tggtaaccat taacagtccc ctccatcgtc atctttacac acatgtcttt cttgactaag
     3661 ctcgccatgg gactctttct cctctttaat cgagaccaag ctagcactgt acctaggact
     3721 gagctagccg tcaagaattc actggccgtc gttttacggt ctcagtcgac tactctagaa
     3781 atattttatc tgattaataa gatgatcttc ttgagatcgt tttggtctgc gcgtaatctc
     3841 ttgctctgaa aacgaaaaaa ccgccttgca gggcggtttt tcgaaggttc tctgagctac
     3901 caactctttg aaccgaggta actggcttgg aggagcgcag tcaccaaaac ttgtcctttc
     3961 agtttagcct taaccggcgc atgacttcaa gactaactcc tctaaatcaa ttaccagtgg
     4021 ctgctgccag tggtgctttt gcatgtcttt ccgggttgga ctcaagacga tagttaccgg
     4081 ataaggcgca gcggtcggac tgaacggggg gttcgtgcat acagtccagc ttggagcgaa
     4141 ctgcctaccc ggaactgagt gtcaggcgtg gaatgagaca aacgcggcca taacagcgga
     4201 atgacaccgg taaaccgaaa ggcaggaaca ggagagcgca cgagggagcc gccaggggga
     4261 aacgcctggt atctttatag tcctgtcggg tttcgccacc actgatttga gcgtcagatt
     4321 tcgtgatgct tgtcaggggg gcggagccta tggaaaaacg gctttgccgc ggccctctca
     4381 cttccctgtt aagtatcttc ctggcatctt ccaggaaatc tccgccccgt tcgtaagcca
     4441 tttccgctcg ccgcagtcga acgaccgagc gtagcgagtc agtgagcgag gaagcggaat
     4501 atatcctgta tcacatattc tgctgacgca ccggtgcagc cttttttctc ctgccacatg
     4561 aagcacttca ctgacaccct catcagtgcc aacatagtaa gccagtatac actccgctag
     4621 cgctgatgtc cggcggtgct tttgccgtta cgcaccaccc cgtcagtagc tgaacaggag
     4681 ggacagctaa agctagaaat gtaccaaatt ttacctattt taggatactt ttcatgaagc
     4741 tcttctattt ttccccagtc ctttaatagt ctacctttag agtctcgtaa atagttatct
     4801 ttgtgacagg ggcctctttt atctttttta atgtaactat atgttattcc aacgtcacta
     4861 ttactattat ccaaatcttt tttagcatgc cagtaagaac tttcataact taactctatc
     4921 tttcgacctc tttgatatac aacaataaag ctatagccag tagtaacaac ctgttttact
     4981 tttgttaaat ctattaactt cttatttatt tttttatgtt tttttgaaaa tttaaatatt
     5041 tctatattca ttcctacact tcctcaaatc caaatggtag cttatgattc tcttctggtt
     5101 tcttttctaa tttttttata tttgcaataa aaactctttt tctatctttg atttttttat
     5161 tgtcccaatt cctccaagta tcatcacaaa ccctttcaat atcatgtaaa tgatgatgtc
     5221 taaatattga tctgacataa tacagatcta ggtctagttc atcacttaac acaacttctc
     5281 taagtctttc agatgcttcg attggtatgt aatcctcttt atttttagta tctaaaggct
     5341 tttgcttaaa ttcttcttct gtctctgcta ccttactaac tgtaaacttg atatttgtaa
     5401 tcttacgacc atgttttctg tgatgatcct tgtcatcata ggttacaaaa atatccgata
     5461 attgattaat ctcttctagt gctggtaata ggaacttatt tttaaaattt gaatatctgt
     5521 tgctgtaact tttaggtaaa tcaaaatcat taatcatatc atcgacatac aatacgcaat
     5581 caactatatt agcataccct gcttgttcgc ctaatttgct tttgagaagt aagtataatc
     5641 tgcttgaata cttactttta aatgaaaata gtaactgtct ttctgcttta gtaaagtact
     5701 cttgtagttg tatcatgtgt ggcattaatg accaatgaaa ctcgcaaatt aaagcactgc
     5761 ttttagggtc tgcttcaata tatgcaaacc agttagctat cttcgtttgt tctttattca
     5821 gccatactgg cttagacatt attgagtgca ttaattgctt caatctcact ctgttatgct
     5881 taacccctgt agctttttca agatcagata ggcttatctt atacctgtga aactctttat
     5941 cttctctttt aaccattgag gcaactaaga atattaagtt ttgttcttct tttgtaaggc
     6001 tatactttcc tgcaacaaga gtattagaca tagctatttc tttgccagca tttacatttt
     6061 taacttcttt catagaacta gagtcattat ctcgatatac aaattctata aaacttctat
     6121 tagtaaaaca actacttcat aaaaaaaagt agttttaacg atacaaaaag tagttttaaa
     6181 ttcaaaaagt gatacaaaaa gtagttttaa attcaaaaag tgatacaaaa agtagtttta
     6241 aattttttaa aaaagtgctt caaagcctta tgtagcaata cttacagagg att
//
