LOCUS       sequence-464935-        6329 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 3736 to 3770)
  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 6329)
  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: pKEK3200
FEATURES             Location/Qualifiers
     source          1..6329
                     /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             3712..3723
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3736..3770)
                     /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(3777..3793)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3847..4392
                     /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     4293..4312
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4509..4526
                     /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 attagctctt atacagctcg
     3001 tccataccca ggctcaagcc cgcagcggtc acaaattcca ccaggaccat gtgatcacgg
     3061 cgttcctgcg gatctttgct cagcacgcta ttggtcgtca gatagtgttg ttccggcagc
     3121 aacaccggac cctcaccaat tggggtgttc tgttggtaat ggtcggccag ttggacgcta
     3181 ccgtcatcaa cattgtggcg aatacggaat tgggcctcca agccgttacg attcttgtct
     3241 gccgtaatgt aaacataatg gctgttgaag ctatactcca gtttatggcc cagaatgtta
     3301 ccatcctctt taaactccaa acccttcaaa tcgatacgat tgaccagcgc ttcaccttca
     3361 aacttaacct ctgcgcgagt tttgtacgtg ccatcttcct tgaaggaaat ggtacgctcc
     3421 tggacgtaac cgtccggcat ggcactttta aagaaatcgt tttggcgcat gtgctccgga
     3481 tacttcgcaa aacactgcac gccatacgac aaggtcgtca ccaaggtcgg ccaaggcacc
     3541 ggcaggcgac cggtcgtgca gatgaactta atttccagtt tgccgttgct cgcgtcgcca
     3601 taaccttcac cgcgaacgct aaaacgatga ccgttcacgt cgccctccag ctcgatcaga
     3661 atcggaacca agcccgagaa aatatcttcg ccactgctca ccatgggact ctttctcctc
     3721 tttaatcgag accaagctag cactgtacct aggactgagc tagccgtcaa gaattcactg
     3781 gccgtcgttt tacggtctca gtcgactact ctagaaatat tttatctgat taataagatg
     3841 atcttcttga gatcgttttg gtctgcgcgt aatctcttgc tctgaaaacg aaaaaaccgc
     3901 cttgcagggc ggtttttcga aggttctctg agctaccaac tctttgaacc gaggtaactg
     3961 gcttggagga gcgcagtcac caaaacttgt cctttcagtt tagccttaac cggcgcatga
     4021 cttcaagact aactcctcta aatcaattac cagtggctgc tgccagtggt gcttttgcat
     4081 gtctttccgg gttggactca agacgatagt taccggataa ggcgcagcgg tcggactgaa
     4141 cggggggttc gtgcatacag tccagcttgg agcgaactgc ctacccggaa ctgagtgtca
     4201 ggcgtggaat gagacaaacg cggccataac agcggaatga caccggtaaa ccgaaaggca
     4261 ggaacaggag agcgcacgag ggagccgcca gggggaaacg cctggtatct ttatagtcct
     4321 gtcgggtttc gccaccactg atttgagcgt cagatttcgt gatgcttgtc aggggggcgg
     4381 agcctatgga aaaacggctt tgccgcggcc ctctcacttc cctgttaagt atcttcctgg
     4441 catcttccag gaaatctccg ccccgttcgt aagccatttc cgctcgccgc agtcgaacga
     4501 ccgagcgtag cgagtcagtg agcgaggaag cggaatatat cctgtatcac atattctgct
     4561 gacgcaccgg tgcagccttt tttctcctgc cacatgaagc acttcactga caccctcatc
     4621 agtgccaaca tagtaagcca gtatacactc cgctagcgct gatgtccggc ggtgcttttg
     4681 ccgttacgca ccaccccgtc agtagctgaa caggagggac agctaaagct agaaatgtac
     4741 caaattttac ctattttagg atacttttca tgaagctctt ctatttttcc ccagtccttt
     4801 aatagtctac ctttagagtc tcgtaaatag ttatctttgt gacaggggcc tcttttatct
     4861 tttttaatgt aactatatgt tattccaacg tcactattac tattatccaa atctttttta
     4921 gcatgccagt aagaactttc ataacttaac tctatctttc gacctctttg atatacaaca
     4981 ataaagctat agccagtagt aacaacctgt tttacttttg ttaaatctat taacttctta
     5041 tttatttttt tatgtttttt tgaaaattta aatatttcta tattcattcc tacacttcct
     5101 caaatccaaa tggtagctta tgattctctt ctggtttctt ttctaatttt tttatatttg
     5161 caataaaaac tctttttcta tctttgattt ttttattgtc ccaattcctc caagtatcat
     5221 cacaaaccct ttcaatatca tgtaaatgat gatgtctaaa tattgatctg acataataca
     5281 gatctaggtc tagttcatca cttaacacaa cttctctaag tctttcagat gcttcgattg
     5341 gtatgtaatc ctctttattt ttagtatcta aaggcttttg cttaaattct tcttctgtct
     5401 ctgctacctt actaactgta aacttgatat ttgtaatctt acgaccatgt tttctgtgat
     5461 gatccttgtc atcataggtt acaaaaatat ccgataattg attaatctct tctagtgctg
     5521 gtaataggaa cttattttta aaatttgaat atctgttgct gtaactttta ggtaaatcaa
     5581 aatcattaat catatcatcg acatacaata cgcaatcaac tatattagca taccctgctt
     5641 gttcgcctaa tttgcttttg agaagtaagt ataatctgct tgaatactta cttttaaatg
     5701 aaaatagtaa ctgtctttct gctttagtaa agtactcttg tagttgtatc atgtgtggca
     5761 ttaatgacca atgaaactcg caaattaaag cactgctttt agggtctgct tcaatatatg
     5821 caaaccagtt agctatcttc gtttgttctt tattcagcca tactggctta gacattattg
     5881 agtgcattaa ttgcttcaat ctcactctgt tatgcttaac ccctgtagct ttttcaagat
     5941 cagataggct tatcttatac ctgtgaaact ctttatcttc tcttttaacc attgaggcaa
     6001 ctaagaatat taagttttgt tcttcttttg taaggctata ctttcctgca acaagagtat
     6061 tagacatagc tatttctttg ccagcattta catttttaac ttctttcata gaactagagt
     6121 cattatctcg atatacaaat tctataaaac ttctattagt aaaacaacta cttcataaaa
     6181 aaaagtagtt ttaacgatac aaaaagtagt tttaaattca aaaagtgata caaaaagtag
     6241 ttttaaattc aaaaagtgat acaaaaagta gttttaaatt ttttaaaaaa gtgcttcaaa
     6301 gccttatgta gcaatactta cagaggatt
//
