LOCUS       sequence-488064-        3149 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 3149)
  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  JOURNAL   Unpublished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 3149)
  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            "Unpublished"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_Red
FEATURES             Location/Qualifiers
     source          1..3149
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        22..69
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      809..880
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(995..1011)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(995..1011)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1008..1030)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1191..1736)
                     /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(1271..1290)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1828..1852)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1952..2743,2744..2812))
                     /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     2575..2594
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2813..2917)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3016..3038
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3030..3047
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3031..3047
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctcagga gagagacgcc tgttgacaat taatcatcgg catagtatat cggcatagta
       61 taatacgacc aaagaaacaa aatgaggagg tactgaaatg gtgagcaaag gcgaagcggt
      121 gattaaagaa tttatgcgct ttaaagtgca tatggaaggc agcatgaacg gccatgaatt
      181 tgaaattgaa ggcgaaggcg aaggccgccc gtatgaaggc acccagaccg cgaaactgaa
      241 agtgaccaaa ggcggcccgc tgccgtttag ctgggatatt ctgagcccgc agtttatgta
      301 tggcagccgc gcgtttatta aacatccggc ggatattccg gattattata aacagagctt
      361 tccggaaggc tttaaatggg aacgcgtgat gaactttgaa gatggcggcg cggtgaccgt
      421 gacccaggat accagcctgg aagatggcac cctgatttat aaagtgaaac tgcgcggcac
      481 caactttccg ccggatggcc cggtgatgca gaaaaaaacc atgggctggg aagcgagcac
      541 cgaacgcctg tatccggaag atggcgtgct gaaaggcgat attaaaatgg cgctgcgcct
      601 gaaagatggc ggccgctatc tggcggattt taaaaccacc tataaagcga aaaaaccggt
      661 gcagatgccg ggcgcgtata acgtggatcg caaactggat attaccagcc ataacgaaga
      721 ttataccgtg gtggaacagt atgaacgcag cgaaggccgc catagcaccg gcggcatgga
      781 tgaactgtat aaataagctt ccaggcatca aataaaacga aaggctcagt cgaaagactg
      841 ggcctttcgt tttatctgtt gtttgtcggt gaacgctctc cgtctctcgc tgtcatgaga
      901 ccggatcttg acacggttat ccacagaatc aggggataag gctaggtgga ggctcagtga
      961 tgataagtct gcgatggtgg atgcatgtgt catggtcata gctgtttcct gtgtgaaatt
     1021 gttatccgct cagagggcac aatcctattc cgcgctatcc gacaatctcc aagacattag
     1081 gtggagttca gttcggcgag cggaaatggc ttacgaacgg ggcggagatt tcctggaaga
     1141 tgccaggaag atacttaaca gggaagtgag agggccgcgg caaagccgtt tttccatagg
     1201 ctccgccccc ctgacaagca tcacgaaatc tgacgctcaa atcagtggtg gcgaaacccg
     1261 acaggactat aaagatacca ggcgtttccc cctggcggct ccctcgtgcg ctctcctgtt
     1321 cctgcctttc ggtttaccgg tgtcattccg ctgttatggc cgcgtttgtc tcattccacg
     1381 cctgacactc agttccgggt aggcagttcg ctccaagctg gactgtatgc acgaaccccc
     1441 cgttcagtcc gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggaaag
     1501 acatgcaaaa gcaccactgg cagcagccac tggtaattga tttagaggag ttagtcttga
     1561 agtcatgcgc cggttaaggc taaactgaaa ggacaagttt tggtgactgc gctcctccaa
     1621 gccagttacc tcggttcaaa gagttggtag ctcagagaac cttcgaaaaa ccgccctgca
     1681 aggcggtttt ttcgttttca gagcaagaga ttacgcgcag accaaaacga tctcaagaag
     1741 atcatcttat taagtctgac gctctattca acaaagccgc cgtccatggg tagggggctt
     1801 caaatcgtcc tcgtgatacc aattcggagc ctgctttttt gtacaaactt gttgataatg
     1861 gcaattcaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     1921 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     1981 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     2041 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagagccac
     2101 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     2161 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     2221 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     2281 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     2341 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     2401 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     2461 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     2521 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     2581 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     2641 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     2701 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     2761 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     2821 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     2881 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccag
     2941 atacctgaaa caaaacccat cgtacggcca aggaagtctc caataactgt gatccaccac
     3001 aagcgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtcat gcataatccg
     3061 cacgcatctg gaataaggaa gtgccattcc gcctgacctt ttaacgcgaa ttttaacaaa
     3121 atattaacgc ttacaatttt aacggatcc
//
