LOCUS       sequence-464934-        6338 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 3745 to 3779)
  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 6338)
  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: pKEK3194
FEATURES             Location/Qualifiers
     source          1..6338
                     /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"
     CDS             complement(2985..3002)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3721..3732
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3745..3779)
                     /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(3786..3802)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3856..4401
                     /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     4302..4321
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4518..4535
                     /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 atcagtgatg atgatgatgg
     3001 tgctgatatg tgtctaagtc tacggctttt acaacttcta cgaggcacat gtgatctctt
     3061 ctctcatctc tatctttaga taatttagtg tggtaagtga tatgatgata ccttggtata
     3121 tgcacagcag cacttccggc gagaggcctg ttcatctggg aacacttggt gaccagtttt
     3181 tcagtcaccc cctcaatatc gtatgcttga ttaaattcca ctcggattcc attgttaacg
     3241 gtatcgggca gtatatacag gatgctggga ggacattgaa aagcaacgtt cttcctcaga
     3301 atgtgtccat cctttttgaa gttttctccg gtcaaggtca ccctgttgta tatagaccct
     3361 ctctcatagg ttaccatagc tcttgtctta tatacgccgt cgccttcaaa acttatagtt
     3421 ctttcttggg tgtaaccctc tggcattgct gacttaaaga aatcttttat atggctgggg
     3481 tacttagcaa aacattggac cccatagcta agggttgaca ccagtgtagc ccagggtact
     3541 ggtaaatcgc cagttgtaca gatatatttg gccttaatag taccagtggt agcgtcgcca
     3601 gttccctccc ccttgattat gaacttcatg ccctccacat ccccttcaag ttctgttatg
     3661 tagggtattt ctttttcaaa cagtttagca ccctctgtca aggcagtagc catgggactc
     3721 tttctcctct ttaatcgaga ccaagctagc actgtaccta ggactgagct agccgtcaag
     3781 aattcactgg ccgtcgtttt acggtctcag tcgactactc tagaaatatt ttatctgatt
     3841 aataagatga tcttcttgag atcgttttgg tctgcgcgta atctcttgct ctgaaaacga
     3901 aaaaaccgcc ttgcagggcg gtttttcgaa ggttctctga gctaccaact ctttgaaccg
     3961 aggtaactgg cttggaggag cgcagtcacc aaaacttgtc ctttcagttt agccttaacc
     4021 ggcgcatgac ttcaagacta actcctctaa atcaattacc agtggctgct gccagtggtg
     4081 cttttgcatg tctttccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     4141 cggactgaac ggggggttcg tgcatacagt ccagcttgga gcgaactgcc tacccggaac
     4201 tgagtgtcag gcgtggaatg agacaaacgc ggccataaca gcggaatgac accggtaaac
     4261 cgaaaggcag gaacaggaga gcgcacgagg gagccgccag ggggaaacgc ctggtatctt
     4321 tatagtcctg tcgggtttcg ccaccactga tttgagcgtc agatttcgtg atgcttgtca
     4381 ggggggcgga gcctatggaa aaacggcttt gccgcggccc tctcacttcc ctgttaagta
     4441 tcttcctggc atcttccagg aaatctccgc cccgttcgta agccatttcc gctcgccgca
     4501 gtcgaacgac cgagcgtagc gagtcagtga gcgaggaagc ggaatatatc ctgtatcaca
     4561 tattctgctg acgcaccggt gcagcctttt ttctcctgcc acatgaagca cttcactgac
     4621 accctcatca gtgccaacat agtaagccag tatacactcc gctagcgctg atgtccggcg
     4681 gtgcttttgc cgttacgcac caccccgtca gtagctgaac aggagggaca gctaaagcta
     4741 gaaatgtacc aaattttacc tattttagga tacttttcat gaagctcttc tatttttccc
     4801 cagtccttta atagtctacc tttagagtct cgtaaatagt tatctttgtg acaggggcct
     4861 cttttatctt ttttaatgta actatatgtt attccaacgt cactattact attatccaaa
     4921 tcttttttag catgccagta agaactttca taacttaact ctatctttcg acctctttga
     4981 tatacaacaa taaagctata gccagtagta acaacctgtt ttacttttgt taaatctatt
     5041 aacttcttat ttattttttt atgttttttt gaaaatttaa atatttctat attcattcct
     5101 acacttcctc aaatccaaat ggtagcttat gattctcttc tggtttcttt tctaattttt
     5161 ttatatttgc aataaaaact ctttttctat ctttgatttt tttattgtcc caattcctcc
     5221 aagtatcatc acaaaccctt tcaatatcat gtaaatgatg atgtctaaat attgatctga
     5281 cataatacag atctaggtct agttcatcac ttaacacaac ttctctaagt ctttcagatg
     5341 cttcgattgg tatgtaatcc tctttatttt tagtatctaa aggcttttgc ttaaattctt
     5401 cttctgtctc tgctacctta ctaactgtaa acttgatatt tgtaatctta cgaccatgtt
     5461 ttctgtgatg atccttgtca tcataggtta caaaaatatc cgataattga ttaatctctt
     5521 ctagtgctgg taataggaac ttatttttaa aatttgaata tctgttgctg taacttttag
     5581 gtaaatcaaa atcattaatc atatcatcga catacaatac gcaatcaact atattagcat
     5641 accctgcttg ttcgcctaat ttgcttttga gaagtaagta taatctgctt gaatacttac
     5701 ttttaaatga aaatagtaac tgtctttctg ctttagtaaa gtactcttgt agttgtatca
     5761 tgtgtggcat taatgaccaa tgaaactcgc aaattaaagc actgctttta gggtctgctt
     5821 caatatatgc aaaccagtta gctatcttcg tttgttcttt attcagccat actggcttag
     5881 acattattga gtgcattaat tgcttcaatc tcactctgtt atgcttaacc cctgtagctt
     5941 tttcaagatc agataggctt atcttatacc tgtgaaactc tttatcttct cttttaacca
     6001 ttgaggcaac taagaatatt aagttttgtt cttcttttgt aaggctatac tttcctgcaa
     6061 caagagtatt agacatagct atttctttgc cagcatttac atttttaact tctttcatag
     6121 aactagagtc attatctcga tatacaaatt ctataaaact tctattagta aaacaactac
     6181 ttcataaaaa aaagtagttt taacgataca aaaagtagtt ttaaattcaa aaagtgatac
     6241 aaaaagtagt tttaaattca aaaagtgata caaaaagtag ttttaaattt tttaaaaaag
     6301 tgcttcaaag ccttatgtag caatacttac agaggatt
//
