LOCUS       sequence-493265-        3037 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 3037)
  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  JOURNAL   Unpublished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 3037)
  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            "Unpublished"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: pBSKo2_Red
FEATURES             Location/Qualifiers
     source          1..3037
                     /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"
     rep_origin      complement(996..1584)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1076..1095)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        1699..1801
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     CDS             1802..2461
                     /codon_start=1
                     /gene="cat"
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     primer_bind     complement(1868..1887)
                     /label=CAT-R
                     /note="Chloramphenicol resistance gene, reverse primer"
     rep_origin      2574..3029
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2661..2680)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2871..2892
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
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 agtgacacgg
      901 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
      961 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     1021 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     1081 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     1141 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     1201 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     1261 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     1321 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     1381 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     1441 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     1501 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     1561 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     1621 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     1681 acctagatcc ttttgtgatg atcggcacgt aagaggttcc aactttcacc ataatgaaat
     1741 aagatcacta ccgggcgtat tttttgagtt atcgagattt tcaggagcta aggaagctaa
     1801 aatggagaaa aaaatcactg gatataccac cgttgatata tcccaatggc atcgtaaaga
     1861 acattttgag gcatttcagt cagttgctca atgtacctat aaccagaccg ttcagctgga
     1921 tattacggcc tttttaaaga ccgtaaagaa aaataagcac aagttttatc cggcctttat
     1981 tcacattctt gcccgcctga tgaatgctca tccggaattc cgtatggcaa tgaaagacgg
     2041 tgagctggtg atatgggata gtgttcaccc ttgttacacc gttttccatg agcaaactga
     2101 aacgttttca tcgctctgga gtgaatacca cgacgatttc cggcagtttc tacacatata
     2161 ttcgcaagat gtggcgtgtt acggtgaaaa cctggcctat ttccctaaag ggtttattga
     2221 gaatatgttt ttcgtgtcag ccaatccctg ggtgagtttc accagttttg atttaaacgt
     2281 ggccaatatg gacaacttct tcgcccccgt tttcaccatg ggcaaatatt atacgcaagg
     2341 cgacaaggtg ctgatgccgc tggcgattca ggttcatcat gccgtttgtg atggcttcca
     2401 tgtcggcaga atgcttaatg aattacaaca gtactgcgat gagtggcagg gcggggcgta
     2461 atttttttaa ggcagttatt ggtgccctta aacgcctggt tgctacgcct gaataagtga
     2521 taataagcgg atgaatggca gaaattcgat gataagctgt caaacatgat gttacgcgcc
     2581 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     2641 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     2701 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     2761 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccagcgcc
     2821 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     2881 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     2941 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     3001 ttttaacaaa atattaacgc ttacaatttt aactgca
//
