LOCUS       sequence-488062-        2750 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  GoldenBraid2.0 expression vector..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2750)
  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  JOURNAL   Unpublished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 2750)
  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            "Unpublished"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: p15Ao1_Blue
FEATURES             Location/Qualifiers
     source          1..2750
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..69
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        123..141
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     123..140
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    148..204
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(206..225)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(207..225)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(232..248)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(457..476)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(596..612)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(596..612)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(609..631)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(792..1337)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A 
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin.
                     "
     primer_bind     complement(872..891)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1429..1453)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1553..2344,2345..2413))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2176..2195
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2414..2518)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2632..2648
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctcagga gagagacgcc tgttgacaat taatcatcgg catagtatat cggcatagta
       61 taatacgaca aggtgaggaa ctaaacccag gaggcagatc atgaccatga ttacgccaag
      121 ctatttaggt gacactatag aatactcaag cttgcatgcc tgcaggtcga ctctagagga
      181 tccccgggta ccgagctcga attcgcccta tagtgagtcg tattacaatt cactggccgt
      241 cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc
      301 acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca
      361 acagttgcgc agcctgaatg gcgaatgacg cctgatgcgg tattttctcc ttacgcatct
      421 gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg atgccgcata
      481 gcgtctctcg ctgtcatgag accggatctt gacacggtta tccacagaat caggggataa
      541 ggctaggtgg aggctcagtg atgataagtc tgcgatggtg gatgcatgtg tcatggtcat
      601 agctgtttcc tgtgtgaaat tgttatccgc tcagagggca caatcctatt ccgcgctatc
      661 cgacaatctc caagacatta ggtggagttc agttcggcga gcggaaatgg cttacgaacg
      721 gggcggagat ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg
      781 gcaaagccgt ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca
      841 aatcagtggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc
      901 tccctcgtgc gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg
      961 ccgcgtttgt ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct
     1021 ggactgtatg cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg
     1081 tcttgagtcc aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg
     1141 atttagagga gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt
     1201 ttggtgactg cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa
     1261 ccttcgaaaa accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca
     1321 gaccaaaacg atctcaagaa gatcatctta ttaagtctga cgctctattc aacaaagccg
     1381 ccgtccatgg gtagggggct tcaaatcgtc ctcgtgatac caattcggag cctgcttttt
     1441 tgtacaaact tgttgataat ggcaattcaa ggatcttcac ctagatcctt ttaaattaaa
     1501 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     1561 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     1621 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     1681 caatgatacc gcgagagcca cgctcaccgg ctccagattt atcagcaata aaccagccag
     1741 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     1801 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     1861 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     1921 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     1981 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     2041 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     2101 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     2161 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     2221 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     2281 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     2341 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     2401 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     2461 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     2521 catttccccg aaaagtgcca gatacctgaa acaaaaccca tcgtacggcc aaggaagtct
     2581 ccaataactg tgatccacca caagcgccag ggttttccca gtcacgacgt tgtaaaacga
     2641 cggccagtca tgcataatcc gcacgcatct ggaataagga agtgccattc cgcctgacct
     2701 tttaacgcga attttaacaa aatattaacg cttacaattt taacggatcc
//
