LOCUS       sequence-492107-        2806 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 2806)
  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  JOURNAL   Unpublished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 2806)
  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            "Unpublished"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: pR6Ka1_Red
FEATURES             Location/Qualifiers
     source          1..2806
                     /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(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      1034..1106
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      1162..1518
                     /label=R6K gamma ori
                     /note="gamma replication origin from E. coli plasmid R6K; 
                     requires the R6K initiator protein pi for replication"
     CDS             complement(1632..2447)
                     /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(1632..2447)
                     /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     2355..2374
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        complement(2448..2550)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(2569..2596)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     2737..2754
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2738..2754
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2761..2780
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2761..2779
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
ORIGIN
        1 cgtctcagga gagagacccc 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 ctctcggtct ctcgctgtca tgagacgaat tctgacttgt
      901 attccgcggt tctatagtgt cacctaaata gcttggcgta atttaggtca tagctgtttc
      961 ctgtcggacc aaaacgaaaa aaggcccccc tttcgggagg cctcttttct ggaatttggt
     1021 accgagcagg catcaaataa aacgaaaggc tcagtcgaaa gactgggcct ttcgttttat
     1081 ctgttgtttg tcggttgaac gctctctgac gtaccgcggc cgcgtagagg atctgttgat
     1141 cagcagttca acctgttgat agtacgtact aagctctcat gtttcacgta ctaagctctc
     1201 atgtttaacg tactaagctc tcatgtttaa cgaactaaac cctcatggct aacgtactaa
     1261 gctctcatgg ctaacgtact aagctctcat gtttcacgta ctaagctctc atgtttgaac
     1321 aataaaatta atataaatca gcaacttaaa tagcctctaa ggttttaagt tttataagaa
     1381 aaaaaagaat atataaggct tttaaagctt ttaaggttta acggttgtgg acaacaagcc
     1441 agggatgtaa cgcactgaga agcccttaga gcctctcaaa gcaattttga gtgacacagg
     1501 aacacttaac ggctgacatg tgatcatgtt tgacagctta tcatcgaatt tctgccattc
     1561 atccgcttat tatcacttat tcaggcgtag caaccaggcg tttaagggca ccaataactg
     1621 ccttaaaaaa attagaaaaa ctcatcgagc atcaaatgaa actgcaattt attcatatca
     1681 ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg
     1741 aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac tcgtccaaca
     1801 tcaatacaac ctatcaattt cccctcgtca aaaataaggt tatcaagtga gaaatcacca
     1861 tgagtgacta ctgaatccgg tgagaatggc aaaagtttgt gcatttcttt ccagacttgt
     1921 tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa accgttattc
     1981 attcgtgatt gcgcctgagc gagtcgaaat acgcggtcgc tgttaaaagg acaattacaa
     2041 acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat gttttcacct
     2101 gaatcaggat attcttctaa tacctggaat gctgttttcc ctgggatcgc agtggtgagt
     2161 aaccacgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg cataaattcc
     2221 gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct acctttgcca
     2281 tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcggtagat tgtcgcacct
     2341 gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc catgttggaa
     2401 tttaatcgcg gccttgagca agacgtttcc cgttgaatat ggctcatttt agcttcctta
     2461 gctcctgaaa atctcgataa ctcaaaaaat acgcccggta gtgatcttat ttcattatgg
     2521 tgaaagttgg aacctcttac gtgccgatca taacattgta gaaacgcaaa aaggccatcc
     2581 gtcaggatgg ccttctgctt aatttgatgc ctggcagttt atggcgggcg tcctgcccgc
     2641 caccctttca gccaaaaaac ttaagaccgc cggtcttgtc cactaccttg cagtaatgcg
     2701 gtggacagga tcggcggttt tcttttctct tctcaatgta aaacgacggc cagtgaattg
     2761 taatacgact cactataggg ttcgaattct atcggtaagc acaaat
//
