LOCUS       sequence-488067-        2627 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 2627)
  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  JOURNAL   Unpublished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 2627)
  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            "Unpublished"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: pR6Ko2_Red
FEATURES             Location/Qualifiers
     source          1..2627
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..60
                     /label=BBa_J23111
                     /note="Anderson constitutive promoter 
                     (https://parts.igem.org/Promoters/Catalog/Anderson). 
                     Measured strength relative to BBa_J223100: 0.58"
     terminator      798..869
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     promoter        complement(911..929)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(912..929)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(947..963)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(947..963)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     terminator      1029..1101
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1157..1513
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1627..2286)
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     2201..2220
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     promoter        complement(2287..2389)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2408..2435)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2576..2593
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2577..2593
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2600..2619
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2600..2618
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 ggtctcagtc aggagagaga cgcctttgac ggctagctca gtcctaggta tagtgctagc
       61 tcaaagagaa agaggggaca aactgaatgg tgagcaaagg cgaagcggtg attaaagaat
      121 ttatgcgctt taaagtgcat atggaaggca gcatgaacgg ccatgaattt gaaattgaag
      181 gcgaaggcga aggccgcccg tatgaaggca cccagaccgc gaaactgaaa gtgaccaaag
      241 gcggcccgct gccgtttagc tgggatattc tgagcccgca gtttatgtat ggcagccgcg
      301 cgtttattaa acatccggcg gatattccgg attattataa acagagcttt ccggaaggct
      361 ttaaatggga acgcgtgatg aactttgaag atggcggcgc ggtgaccgtg acccaggata
      421 ccagcctgga agatggcacc ctgatttata aagtgaaact gcgcggcacc aactttccgc
      481 cggatggccc ggtgatgcag aaaaaaacca tgggctggga agcgagcacc gaacgcctgt
      541 atccggaaga tggcgtgctg aaaggcgata ttaaaatggc gctgcgcctg aaagatggcg
      601 gccgctatct ggcggatttt aaaaccacct ataaagcgaa aaaaccggtg cagatgccgg
      661 gcgcgtataa cgtggatcgc aaactggata ttaccagcca taacgaagat tataccgtgg
      721 tggaacagta tgaacgcagc gaaggccgcc atagcaccgg cggcatggat gaactgtata
      781 aataagcttc caggcatcaa ataaaacgaa aggctcagtc gaaagactgg gcctttcgtt
      841 ttatctgttg tttgtcggtg aacgctctcc gtctctcgct tgagacctgc agtgacttgt
      901 attccgcggt tctatagtgt cacctaaata gcttggcgta atttaggtca tagctgtttc
      961 ctgtcggacc aaaacgaaaa aaggcccccc tttcgggagg cctctggaat ttggtaccga
     1021 gcaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1081 gtttgtcggt tgaacgctct ctgacgtacc gcggccgcgt agaggatctg ttgatcagca
     1141 gttcaacctg ttgatagtac gtactaagct ctcatgtttc acgtactaag ctctcatgtt
     1201 taacgtacta agctctcatg tttaacgaac taaaccctca tggctaacgt actaagctct
     1261 catggctaac gtactaagct ctcatgtttc acgtactaag ctctcatgtt tgaacaataa
     1321 aattaatata aatcagcaac ttaaatagcc tctaaggttt taagttttat aagaaaaaaa
     1381 agaatatata aggcttttaa agcttttaag gtttaacggt tgtggacaac aagccaggga
     1441 tgtaacgcac tgagaagccc ttagagcctc tcaaagcaat tttgagtgac acaggaacac
     1501 ttaacggctg acatgtgatc atgtttgaca gcttatcatc gaatttctgc cattcatccg
     1561 cttattatca cttattcagg cgtagcaacc aggcgtttaa gggcaccaat aactgcctta
     1621 aaaaaattac gccccgccct gccactcgtc gcaatactgt tgtaattcat taagcattct
     1681 gccgacatgg aagccatcac aaacggcatg atgaacctga atcgccagcg gcatcagcac
     1741 cttgtcgcct tgcgtatagt atttgcccat cgtgaaaacg ggggcgaaga agttgtccat
     1801 attggccacg tttaaatcaa aactggtgaa actcacccag ggattggctg acacgaaaaa
     1861 catattctca ataaaccctt tagggaaata ggccaggttt tcaccgtaac acgccacatc
     1921 ttgcgaatat atgtgtagaa actgccggaa atcgtcgtgg tattcactcc agaggcttga
     1981 aaacgtttca gtttgctcat ggaaaacggt gtaacaaggg tgaacactat cccatatcac
     2041 cagctcaccg tctttcattg ccatacgaaa ttccggatga gcattcatca ggcgggcaag
     2101 aatgtgaata aaggccggat aaaacttgtg cttatttttc tttacggtct ttaaaaaggc
     2161 cgtaatatcc agttgaacgg tctggttata ggtacattga gcaactgact gaaatgcctc
     2221 aaaatgttct ttacgatgcc attgggatat atcaacggtg gtatatccag tgattttttt
     2281 ctccatttta gcttccttag ctcctgaaaa tctcgataac tcaaaaaata cgcccggtag
     2341 tgatcttatt tcattatggt gaaagttgga acctcttacg tgccgatcat aacattgtag
     2401 aaacgcaaaa aggccatccg tcaggatggc cttctgctta atttgatgcc tggcagttta
     2461 tggcgggcgt cctgcccgcc accctttcag ccaaaaaact taagaccgcc ggtcttgtcc
     2521 actaccttgc agtaatgcgg tggacaggat cggcggtttt cttttctctt ctcaatgtaa
     2581 aacgacggcc agtgaattgt aatacgactc actatagggt tcctgca
//
