LOCUS       sequence-492930-        2747 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 2747)
  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  JOURNAL   Unpublished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 2747)
  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            "Unpublished"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: p15Ao2_Blue
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..73
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        127..145
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     127..144
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     misc_feature    152..208
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(210..229)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(211..229)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(236..252)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(461..480)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(595..611)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(595..611)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(608..630)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(791..1336)
                     /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(871..890)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1428..1452)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1552..2343,2344..2412))
                     /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     2175..2194
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2413..2517)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     2631..2647
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctcagtc aggagagaga cgcctgttga caattaatca tcggcatagt atatcggcat
       61 agtataatac gacaaggtga ggaactaaac ccaggaggca gatcatgacc atgattacgc
      121 caagctattt aggtgacact atagaatact caagcttgca tgcctgcagg tcgactctag
      181 aggatccccg ggtaccgagc tcgaattcgc cctatagtga gtcgtattac aattcactgg
      241 ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg
      301 cagcacaccc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt
      361 cccaacagtt gcgcagcctg aatggcgaat gacgcctgat gcggtatttt ctccttacgc
      421 atctgtgcgg tatttcacac cgcatatggt gcactctcag tacaatctgc tctgatgccg
      481 catagcgtct ctcgcttgag acctgcagtg acacggttat ccacagaatc aggggataag
      541 gctaggtgga ggctcagtga tgataagtct gcgatggtgg atgcatgtgt catggtcata
      601 gctgtttcct gtgtgaaatt gttatccgct cagagggcac aatcctattc cgcgctatcc
      661 gacaatctcc aagacattag gtggagttca gttcggcgag cggaaatggc ttacgaacgg
      721 ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag agggccgcgg
      781 caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa
      841 atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggcggct
      901 ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg ctgttatggc
      961 cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg ctccaagctg
     1021 gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg taactatcgt
     1081 cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac tggtaattga
     1141 tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa ggacaagttt
     1201 tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag ctcagagaac
     1261 cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga ttacgcgcag
     1321 accaaaacga tctcaagaag atcatcttat taagtctgac gctctattca acaaagccgc
     1381 cgtccatggg tagggggctt caaatcgtcc tcgtgatacc aattcggagc ctgctttttt
     1441 gtacaaactt gttgataatg gcaattcaag gatcttcacc tagatccttt taaattaaaa
     1501 atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg
     1561 cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg
     1621 actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc
     1681 aatgataccg cgagagccac gctcaccggc tccagattta tcagcaataa accagccagc
     1741 cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa
     1801 ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc
     1861 cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg
     1921 ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc
     1981 cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat
     2041 ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg
     2101 tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc
     2161 ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg
     2221 aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat
     2281 gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg
     2341 gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg
     2401 ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct
     2461 catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac
     2521 atttccccga aaagtgccag atacctgaaa caaaacccat cgtacggcca aggaagtctc
     2581 caataactgt gatccaccac aagcgccagg gttttcccag tcacgacgtt gtaaaacgac
     2641 ggccagtcat gcataatccg cacgcatctg gaataaggaa gtgccattcc gcctgacctt
     2701 ttaacgcgaa ttttaacaaa atattaacgc ttacaatttt aactgca
//
