LOCUS       sequence-464932-        6311 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 3718 to 3752)
  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 6311)
  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: pKEK3198
FEATURES             Location/Qualifiers
     source          1..6311
                     /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(2982..2999)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     RBS             3694..3705
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     promoter        complement(3718..3752)
                     /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(3759..3775)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      3829..4374
                     /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     4275..4294
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4491..4508
                     /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 aatggtgatg gtgatggtgg
     3001 ccaagcttag aaggtgcggc ctcgcagtac ctggccacgg ctgattcata ttgttcgtag
     3061 aacttccctt gttcggcctt tagaatttct atacgatggt ccccgaaatg aaattctggc
     3121 atattaacag cgttggatgg ctttcggctc ctgtaggtag ttcttaagtg tgaagttagg
     3181 tggttcccgt cggtacattt taaggccatt agactctgac cgcaaagtac tccgtccctg
     3241 gggtagagca tctccacgca tggttcccag ccagctgtct ttttctgcat aactggccca
     3301 ttggcgggga aattagcgcc gatcacattt actttgaaga taaaattgtt gccctgtaga
     3361 gaggtatcct gatgtatggt gagataacca ccgtcctcaa aaatctgagt cctttcccaa
     3421 gtgaagccct ctggaaagct ttctttgaaa taatcgggaa taccacttac gtgcttgatg
     3481 aaagtttttg agccatacat acaacatgga gctagaatgt cgtaagcaaa tgggagaggt
     3541 ccaccctcag tgattctgat cttctcgacc tgagtaccct caaatggctt accctcacct
     3601 tcgcctacgc atttgaagtg gtgaccatta acggttccct ccatagtcat tttgatgcac
     3661 atatccttct ttactaggct ggccatggga ctctttctcc tctttaatcg agaccaagct
     3721 agcactgtac ctaggactga gctagccgtc aagaattcac tggccgtcgt tttacggtct
     3781 cagtcgacta ctctagaaat attttatctg attaataaga tgatcttctt gagatcgttt
     3841 tggtctgcgc gtaatctctt gctctgaaaa cgaaaaaacc gccttgcagg gcggtttttc
     3901 gaaggttctc tgagctacca actctttgaa ccgaggtaac tggcttggag gagcgcagtc
     3961 accaaaactt gtcctttcag tttagcctta accggcgcat gacttcaaga ctaactcctc
     4021 taaatcaatt accagtggct gctgccagtg gtgcttttgc atgtctttcc gggttggact
     4081 caagacgata gttaccggat aaggcgcagc ggtcggactg aacggggggt tcgtgcatac
     4141 agtccagctt ggagcgaact gcctacccgg aactgagtgt caggcgtgga atgagacaaa
     4201 cgcggccata acagcggaat gacaccggta aaccgaaagg caggaacagg agagcgcacg
     4261 agggagccgc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccaccac
     4321 tgatttgagc gtcagatttc gtgatgcttg tcaggggggc ggagcctatg gaaaaacggc
     4381 tttgccgcgg ccctctcact tccctgttaa gtatcttcct ggcatcttcc aggaaatctc
     4441 cgccccgttc gtaagccatt tccgctcgcc gcagtcgaac gaccgagcgt agcgagtcag
     4501 tgagcgagga agcggaatat atcctgtatc acatattctg ctgacgcacc ggtgcagcct
     4561 tttttctcct gccacatgaa gcacttcact gacaccctca tcagtgccaa catagtaagc
     4621 cagtatacac tccgctagcg ctgatgtccg gcggtgcttt tgccgttacg caccaccccg
     4681 tcagtagctg aacaggaggg acagctaaag ctagaaatgt accaaatttt acctatttta
     4741 ggatactttt catgaagctc ttctattttt ccccagtcct ttaatagtct acctttagag
     4801 tctcgtaaat agttatcttt gtgacagggg cctcttttat cttttttaat gtaactatat
     4861 gttattccaa cgtcactatt actattatcc aaatcttttt tagcatgcca gtaagaactt
     4921 tcataactta actctatctt tcgacctctt tgatatacaa caataaagct atagccagta
     4981 gtaacaacct gttttacttt tgttaaatct attaacttct tatttatttt tttatgtttt
     5041 tttgaaaatt taaatatttc tatattcatt cctacacttc ctcaaatcca aatggtagct
     5101 tatgattctc ttctggtttc ttttctaatt tttttatatt tgcaataaaa actctttttc
     5161 tatctttgat ttttttattg tcccaattcc tccaagtatc atcacaaacc ctttcaatat
     5221 catgtaaatg atgatgtcta aatattgatc tgacataata cagatctagg tctagttcat
     5281 cacttaacac aacttctcta agtctttcag atgcttcgat tggtatgtaa tcctctttat
     5341 ttttagtatc taaaggcttt tgcttaaatt cttcttctgt ctctgctacc ttactaactg
     5401 taaacttgat atttgtaatc ttacgaccat gttttctgtg atgatccttg tcatcatagg
     5461 ttacaaaaat atccgataat tgattaatct cttctagtgc tggtaatagg aacttatttt
     5521 taaaatttga atatctgttg ctgtaacttt taggtaaatc aaaatcatta atcatatcat
     5581 cgacatacaa tacgcaatca actatattag cataccctgc ttgttcgcct aatttgcttt
     5641 tgagaagtaa gtataatctg cttgaatact tacttttaaa tgaaaatagt aactgtcttt
     5701 ctgctttagt aaagtactct tgtagttgta tcatgtgtgg cattaatgac caatgaaact
     5761 cgcaaattaa agcactgctt ttagggtctg cttcaatata tgcaaaccag ttagctatct
     5821 tcgtttgttc tttattcagc catactggct tagacattat tgagtgcatt aattgcttca
     5881 atctcactct gttatgctta acccctgtag ctttttcaag atcagatagg cttatcttat
     5941 acctgtgaaa ctctttatct tctcttttaa ccattgaggc aactaagaat attaagtttt
     6001 gttcttcttt tgtaaggcta tactttcctg caacaagagt attagacata gctatttctt
     6061 tgccagcatt tacattttta acttctttca tagaactaga gtcattatct cgatatacaa
     6121 attctataaa acttctatta gtaaaacaac tacttcataa aaaaaagtag ttttaacgat
     6181 acaaaaagta gttttaaatt caaaaagtga tacaaaaagt agttttaaat tcaaaaagtg
     6241 atacaaaaag tagttttaaa ttttttaaaa aagtgcttca aagccttatg tagcaatact
     6301 tacagaggat t
//
