LOCUS       sequence-493266-        2634 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 Cloning vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2634)
  AUTHORS   Cooke MB, Welch KT, Deus Ramirez L, Wen AX, Marciano DC,, Herman C
  TITLE     GoldenBraid2.0 E. coli: A comprehensive and characterized toolkit 
            for enterics.
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 2634)
  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: 
            "Unpublished"
COMMENT     Sequence Label: pR6Ko1_Red
FEATURES             Location/Qualifiers
     source          1..2634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..56
                     /label=BBa_J23111
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.58"
     terminator      794..865
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(912..930)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(913..930)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(948..964)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(948..964)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1035..1107
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1163..1519
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1633..2292)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2207..2226
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2293..2395)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2414..2441)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2582..2599
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2583..2599
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2606..2625
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2606..2624
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcagga gagagacgcc tttgacggct agctcagtcc taggtatagt gctagctcaa
       61 agagaaagag gggacaaact gaatggtgag caaaggcgaa gcggtgatta aagaatttat
      121 gcgctttaaa gtgcatatgg aaggcagcat gaacggccat gaatttgaaa ttgaaggcga
      181 aggcgaaggc cgcccgtatg aaggcaccca gaccgcgaaa ctgaaagtga ccaaaggcgg
      241 cccgctgccg tttagctggg atattctgag cccgcagttt atgtatggca gccgcgcgtt
      301 tattaaacat ccggcggata ttccggatta ttataaacag agctttccgg aaggctttaa
      361 atgggaacgc gtgatgaact ttgaagatgg cggcgcggtg accgtgaccc aggataccag
      421 cctggaagat ggcaccctga tttataaagt gaaactgcgc ggcaccaact ttccgccgga
      481 tggcccggtg atgcagaaaa aaaccatggg ctgggaagcg agcaccgaac gcctgtatcc
      541 ggaagatggc gtgctgaaag gcgatattaa aatggcgctg cgcctgaaag atggcggccg
      601 ctatctggcg gattttaaaa ccacctataa agcgaaaaaa ccggtgcaga tgccgggcgc
      661 gtataacgtg gatcgcaaac tggatattac cagccataac gaagattata ccgtggtgga
      721 acagtatgaa cgcagcgaag gccgccatag caccggcggc atggatgaac tgtataaata
      781 agcttccagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
      841 ctgttgtttg tcggtgaacg ctctccgtct ctcgctgtca tgagaccgga tcctgacttg
      901 tattccgcgg ttctatagtg tcacctaaat agcttggcgt aatttaggtc atagctgttt
      961 cctgtcggac caaaacgaaa aaaggccccc ctttcgggag gcctcttttc tggaatttgg
     1021 taccgagcag gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta
     1081 tctgttgttt gtcggttgaa cgctctctga cgtaccgcgg ccgcgtagag gatctgttga
     1141 tcagcagttc aacctgttga tagtacgtac taagctctca tgtttcacgt actaagctct
     1201 catgtttaac gtactaagct ctcatgttta acgaactaaa ccctcatggc taacgtacta
     1261 agctctcatg gctaacgtac taagctctca tgtttcacgt actaagctct catgtttgaa
     1321 caataaaatt aatataaatc agcaacttaa atagcctcta aggttttaag ttttataaga
     1381 aaaaaaagaa tatataaggc ttttaaagct tttaaggttt aacggttgtg gacaacaagc
     1441 cagggatgta acgcactgag aagcccttag agcctctcaa agcaattttg agtgacacag
     1501 gaacacttaa cggctgacat gtgatcatgt ttgacagctt atcatcgaat ttctgccatt
     1561 catccgctta ttatcactta ttcaggcgta gcaaccaggc gtttaagggc accaataact
     1621 gccttaaaaa aattacgccc cgccctgcca ctcgtcgcaa tactgttgta attcattaag
     1681 cattctgccg acatggaagc catcacaaac ggcatgatga acctgaatcg ccagcggcat
     1741 cagcaccttg tcgccttgcg tatagtattt gcccatcgtg aaaacggggg cgaagaagtt
     1801 gtccatattg gccacgttta aatcaaaact ggtgaaactc acccagggat tggctgacac
     1861 gaaaaacata ttctcaataa accctttagg gaaataggcc aggttttcac cgtaacacgc
     1921 cacatcttgc gaatatatgt gtagaaactg ccggaaatcg tcgtggtatt cactccagag
     1981 gcttgaaaac gtttcagttt gctcatggaa aacggtgtaa caagggtgaa cactatccca
     2041 tatcaccagc tcaccgtctt tcattgccat acgaaattcc ggatgagcat tcatcaggcg
     2101 ggcaagaatg tgaataaagg ccggataaaa cttgtgctta tttttcttta cggtctttaa
     2161 aaaggccgta atatccagtt gaacggtctg gttataggta cattgagcaa ctgactgaaa
     2221 tgcctcaaaa tgttctttac gatgccattg ggatatatca acggtggtat atccagtgat
     2281 ttttttctcc attttagctt ccttagctcc tgaaaatctc gataactcaa aaaatacgcc
     2341 cggtagtgat cttatttcat tatggtgaaa gttggaacct cttacgtgcc gatcataaca
     2401 ttgtagaaac gcaaaaaggc catccgtcag gatggccttc tgcttaattt gatgcctggc
     2461 agtttatggc gggcgtcctg cccgccaccc tttcagccaa aaaacttaag accgccggtc
     2521 ttgtccacta ccttgcagta atgcggtgga caggatcggc ggttttcttt tctcttctca
     2581 atgtaaaacg acggccagtg aattgtaata cgactcacta tagggttcgg atcc
//
