LOCUS       sequence-464928-        6308 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 3715 to 3749)
  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 6308)
  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: pKEK3197
FEATURES             Location/Qualifiers
     source          1..6308
                     /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             3691..3702
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3715..3749)
                     /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(3756..3772)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3826..4371
                     /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     4272..4291
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4488..4505
                     /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 atcaatggtg atggtggtga
     3001 tgtgtttcct tgacatcgac tgtgaagtgt ttagcaactg cgtgttcctg gatttgaaat
     3061 ccatcagcta cttctttctt caccaggacg tgttcaataa tgtggttggg gggcatcttt
     3121 acggccttgt gggccttgta tgatgttgta aattggcatt taagtctatg tcctccttcc
     3181 aataataaga acatggtaac gtcccccttc agtatcccat cacaagcagt gataagttct
     3241 gaactgggtt cccagttaag tgtcttcttc tgcattacag gtccattagc tgggaagttg
     3301 actccgtgaa aggtggaatt gtggataaaa caatcttttt ccagacttat gtgagcagag
     3361 gcggtcccgc agccgccatc ctcgaacatt aatgtacgct cccaagaata gccctcgggc
     3421 aggctttgtt taaagtaatc tggcatatct tcggggtaat cgcaaaaaca tcggttgcca
     3481 taggtgaagg tagcactcag tatatcaaag gcaaaaggga gaggaccccc ctttgtgact
     3541 ctgagtttca gggttttttt accctcgtaa ggtttgcctg caccttcacc ttcaatggta
     3601 aattgatgac cattaacgca accgtccatg tggaatcgca ttctgacatc tcttggcaaa
     3661 acttgttttg acagggaagc catgggactc tttctcctct ttaatcgaga ccaagctagc
     3721 actgtaccta ggactgagct agccgtcaag aattcactgg ccgtcgtttt acggtctcag
     3781 tcgactactc tagaaatatt ttatctgatt aataagatga tcttcttgag atcgttttgg
     3841 tctgcgcgta atctcttgct ctgaaaacga aaaaaccgcc ttgcagggcg gtttttcgaa
     3901 ggttctctga gctaccaact ctttgaaccg aggtaactgg cttggaggag cgcagtcacc
     3961 aaaacttgtc ctttcagttt agccttaacc ggcgcatgac ttcaagacta actcctctaa
     4021 atcaattacc agtggctgct gccagtggtg cttttgcatg tctttccggg ttggactcaa
     4081 gacgatagtt accggataag gcgcagcggt cggactgaac ggggggttcg tgcatacagt
     4141 ccagcttgga gcgaactgcc tacccggaac tgagtgtcag gcgtggaatg agacaaacgc
     4201 ggccataaca gcggaatgac accggtaaac cgaaaggcag gaacaggaga gcgcacgagg
     4261 gagccgccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccaccactga
     4321 tttgagcgtc agatttcgtg atgcttgtca ggggggcgga gcctatggaa aaacggcttt
     4381 gccgcggccc tctcacttcc ctgttaagta tcttcctggc atcttccagg aaatctccgc
     4441 cccgttcgta agccatttcc gctcgccgca gtcgaacgac cgagcgtagc gagtcagtga
     4501 gcgaggaagc ggaatatatc ctgtatcaca tattctgctg acgcaccggt gcagcctttt
     4561 ttctcctgcc acatgaagca cttcactgac accctcatca gtgccaacat agtaagccag
     4621 tatacactcc gctagcgctg atgtccggcg gtgcttttgc cgttacgcac caccccgtca
     4681 gtagctgaac aggagggaca gctaaagcta gaaatgtacc aaattttacc tattttagga
     4741 tacttttcat gaagctcttc tatttttccc cagtccttta atagtctacc tttagagtct
     4801 cgtaaatagt tatctttgtg acaggggcct cttttatctt ttttaatgta actatatgtt
     4861 attccaacgt cactattact attatccaaa tcttttttag catgccagta agaactttca
     4921 taacttaact ctatctttcg acctctttga tatacaacaa taaagctata gccagtagta
     4981 acaacctgtt ttacttttgt taaatctatt aacttcttat ttattttttt atgttttttt
     5041 gaaaatttaa atatttctat attcattcct acacttcctc aaatccaaat ggtagcttat
     5101 gattctcttc tggtttcttt tctaattttt ttatatttgc aataaaaact ctttttctat
     5161 ctttgatttt tttattgtcc caattcctcc aagtatcatc acaaaccctt tcaatatcat
     5221 gtaaatgatg atgtctaaat attgatctga cataatacag atctaggtct agttcatcac
     5281 ttaacacaac ttctctaagt ctttcagatg cttcgattgg tatgtaatcc tctttatttt
     5341 tagtatctaa aggcttttgc ttaaattctt cttctgtctc tgctacctta ctaactgtaa
     5401 acttgatatt tgtaatctta cgaccatgtt ttctgtgatg atccttgtca tcataggtta
     5461 caaaaatatc cgataattga ttaatctctt ctagtgctgg taataggaac ttatttttaa
     5521 aatttgaata tctgttgctg taacttttag gtaaatcaaa atcattaatc atatcatcga
     5581 catacaatac gcaatcaact atattagcat accctgcttg ttcgcctaat ttgcttttga
     5641 gaagtaagta taatctgctt gaatacttac ttttaaatga aaatagtaac tgtctttctg
     5701 ctttagtaaa gtactcttgt agttgtatca tgtgtggcat taatgaccaa tgaaactcgc
     5761 aaattaaagc actgctttta gggtctgctt caatatatgc aaaccagtta gctatcttcg
     5821 tttgttcttt attcagccat actggcttag acattattga gtgcattaat tgcttcaatc
     5881 tcactctgtt atgcttaacc cctgtagctt tttcaagatc agataggctt atcttatacc
     5941 tgtgaaactc tttatcttct cttttaacca ttgaggcaac taagaatatt aagttttgtt
     6001 cttcttttgt aaggctatac tttcctgcaa caagagtatt agacatagct atttctttgc
     6061 cagcatttac atttttaact tctttcatag aactagagtc attatctcga tatacaaatt
     6121 ctataaaact tctattagta aaacaactac ttcataaaaa aaagtagttt taacgataca
     6181 aaaagtagtt ttaaattcaa aaagtgatac aaaaagtagt tttaaattca aaaagtgata
     6241 caaaaagtag ttttaaattt tttaaaaaag tgcttcaaag ccttatgtag caatacttac
     6301 agaggatt
//
