LOCUS       sequence-488065-        3146 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 3146)
  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  JOURNAL   Unpublished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 3146)
  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            "Unpublished"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_Red
FEATURES             Location/Qualifiers
     source          1..3146
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        26..73
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     terminator      813..884
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     complement(994..1010)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(994..1010)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1007..1029)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     rep_origin      complement(1190..1735)
                     /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(1270..1289)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    complement(1827..1851)
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             complement(join(1951..2742,2743..2811))
                     /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     2574..2593
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2812..2916)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3015..3037
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     3029..3046
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     3030..3046
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
ORIGIN
        1 ggtctcagtc aggagagaga cgcctgttga caattaatca tcggcatagt atatcggcat
       61 agtataatac gaccaaagaa acaaaatgag gaggtactga aatggtgagc aaaggcgaag
      121 cggtgattaa agaatttatg cgctttaaag tgcatatgga aggcagcatg aacggccatg
      181 aatttgaaat tgaaggcgaa ggcgaaggcc gcccgtatga aggcacccag accgcgaaac
      241 tgaaagtgac caaaggcggc ccgctgccgt ttagctggga tattctgagc ccgcagttta
      301 tgtatggcag ccgcgcgttt attaaacatc cggcggatat tccggattat tataaacaga
      361 gctttccgga aggctttaaa tgggaacgcg tgatgaactt tgaagatggc ggcgcggtga
      421 ccgtgaccca ggataccagc ctggaagatg gcaccctgat ttataaagtg aaactgcgcg
      481 gcaccaactt tccgccggat ggcccggtga tgcagaaaaa aaccatgggc tgggaagcga
      541 gcaccgaacg cctgtatccg gaagatggcg tgctgaaagg cgatattaaa atggcgctgc
      601 gcctgaaaga tggcggccgc tatctggcgg attttaaaac cacctataaa gcgaaaaaac
      661 cggtgcagat gccgggcgcg tataacgtgg atcgcaaact ggatattacc agccataacg
      721 aagattatac cgtggtggaa cagtatgaac gcagcgaagg ccgccatagc accggcggca
      781 tggatgaact gtataaataa gcttccaggc atcaaataaa acgaaaggct cagtcgaaag
      841 actgggcctt tcgttttatc tgttgtttgt cggtgaacgc tctccgtctc tcgcttgaga
      901 cctgcagtga cacggttatc cacagaatca ggggataagg ctaggtggag gctcagtgat
      961 gataagtctg cgatggtgga tgcatgtgtc atggtcatag ctgtttcctg tgtgaaattg
     1021 ttatccgctc agagggcaca atcctattcc gcgctatccg acaatctcca agacattagg
     1081 tggagttcag ttcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
     1141 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
     1201 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
     1261 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
     1321 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
     1381 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
     1441 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
     1501 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
     1561 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     1621 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     1681 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     1741 tcatcttatt aagtctgacg ctctattcaa caaagccgcc gtccatgggt agggggcttc
     1801 aaatcgtcct cgtgatacca attcggagcc tgcttttttg tacaaacttg ttgataatgg
     1861 caattcaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     1921 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     1981 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     2041 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagagccacg
     2101 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     2161 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     2221 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     2281 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     2341 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     2401 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     2461 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     2521 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     2581 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     2641 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     2701 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     2761 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     2821 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     2881 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccaga
     2941 tacctgaaac aaaacccatc gtacggccaa ggaagtctcc aataactgtg atccaccaca
     3001 agcgccaggg ttttcccagt cacgacgttg taaaacgacg gccagtcatg cataatccgc
     3061 acgcatctgg aataaggaag tgccattccg cctgaccttt taacgcgaat tttaacaaaa
     3121 tattaacgct tacaatttta actgca
//
