LOCUS       sequence-492109-        2790 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 2790)
  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  JOURNAL   Unpublished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 2790)
  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            "Unpublished"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: pR6Ka2_Red
FEATURES             Location/Qualifiers
     source          1..2790
                     /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(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..2448)
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(1633..2448)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     2356..2375
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2449..2551)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2570..2597)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2738..2755
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2739..2755
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2762..2781
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2762..2780
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgtctcagtc aggagagaga cccctttgac 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 aacgctctcg gtctctcgct tgagacgcag ctgtgacttg
      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 aattagaaaa actcatcgag catcaaatga aactgcaatt tattcatatc
     1681 aggattatca ataccatatt tttgaaaaag ccgtttctgt aatgaaggag aaaactcacc
     1741 gaggcagttc cataggatgg caagatcctg gtatcggtct gcgattccga ctcgtccaac
     1801 atcaatacaa cctatcaatt tcccctcgtc aaaaataagg ttatcaagtg agaaatcacc
     1861 atgagtgact actgaatccg gtgagaatgg caaaagtttg tgcatttctt tccagacttg
     1921 ttcaacaggc cagccattac gctcgtcatc aaaatcactc gcatcaacca aaccgttatt
     1981 cattcgtgat tgcgcctgag cgagtcgaaa tacgcggtcg ctgttaaaag gacaattaca
     2041 aacaggaatc gaatgcaacc ggcgcaggaa cactgccagc gcatcaacaa tgttttcacc
     2101 tgaatcagga tattcttcta atacctggaa tgctgttttc cctgggatcg cagtggtgag
     2161 taaccacgca tcatcaggag tacggataaa atgcttgatg gtcggaagag gcataaattc
     2221 cgtcagccag tttagtctga ccatctcatc tgtaacatca ttggcaacgc tacctttgcc
     2281 atgtttcaga aacaactctg gcgcatcggg cttcccatac aatcggtaga ttgtcgcacc
     2341 tgattgcccg acattatcgc gagcccattt atacccatat aaatcagcat ccatgttgga
     2401 atttaatcgc ggccttgagc aagacgtttc ccgttgaata tggctcattt tagcttcctt
     2461 agctcctgaa aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg
     2521 gtgaaagttg gaacctctta cgtgccgatc ataacattgt agaaacgcaa aaaggccatc
     2581 cgtcaggatg gccttctgct taatttgatg cctggcagtt tatggcgggc gtcctgcccg
     2641 ccaccctttc agccaaaaaa cttaagaccg ccggtcttgt ccactacctt gcagtaatgc
     2701 ggtggacagg atcggcggtt ttcttttctc ttctcaatgt aaaacgacgg ccagtgaatt
     2761 gtaatacgac tcactatagg gttccagctg
//
