LOCUS       sequence-442585-        2773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and p15A origin of 
            replication. Carries A-G CIDAR MoClo overhangs for level 0 or 2 
            construct cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2773)
  AUTHORS   Nielsen JR, Lewis MJ, Huang WE
  TITLE     Construction and Characterization of MoClo-Compatible Vectors for 
            Modular Protein Expression in E. coli.
  JOURNAL   ACS Synth Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564.
  PUBMED    39801078
REFERENCE   2  (bases 1 to 2773)
  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: "ACS Synth 
            Biol. 2025 Jan 12. doi: 10.1021/acssynbio.4c00564."
COMMENT     Sequence Label: DVA_p15A_AG
FEATURES             Location/Qualifiers
     source          1..2773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(244..338)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(371..1162,1163..1231))
                     /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     994..1013
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1232..1336)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1404..1422
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1426..1469)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1472..1493
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1507..1815)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1710..1732
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1724..1741
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1725..1741
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1742..1798
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1811..1827)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1811..1827)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1824..1846)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1835..1851
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1859..1865,1866..1883,1884..1889))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1904..1925
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      2067..2138
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2154..2181
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2224..2241)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2358..2773,1..130))
                     /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(2438..2457)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctgcgctcct ccaagccagt tacctcggtt caaagagttg gtagctcaga gaaccttcga
       61 aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa
      121 acgatctcaa gaagatcatc ttattaatca gataaaatat ttctagattt cagtgcaatt
      181 tatctcttca aatgtagcac ctgaagtcag ccccatacga tataagttgt tactagtgct
      241 tggattctca ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg
      301 gagttctgag gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag
      361 agtcccgcga ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc
      421 gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac
      481 catctggccc cagtgctgca atgataccgc gcgacccacg ctcaccggct ccagatttat
      541 cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg
      601 cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata
      661 gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta
      721 tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt
      781 gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag
      841 tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa
      901 gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc
      961 gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt
     1021 taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc
     1081 tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta
     1141 ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa
     1201 taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca
     1261 tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac
     1321 aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa gaaaccatta
     1381 ttataatgac attaacctat aaaaataggc gtatcacgag gcagaatttc agataaaaaa
     1441 aatccttagc tttcgctaag gatgatttct ggaattcgcg gccgcttcta gagactagtg
     1501 gaggtctcag gagatgtctt ccctgcacca tatgcggtgt gaaataccgc acagatgcgt
     1561 aaggagaaaa taccgcatca ggcgccattc gccattcagg ctgcgcaact gttgggaagg
     1621 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctgcaag
     1681 gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa cgacggccag
     1741 tgaattcgag ctcggtaccc ggggatcctc tagagtcgac ctgcaggcat gcaagcttgg
     1801 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     1861 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     1921 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     1981 attaatgaat cggccaacgc gcgggggtta agaagacgtt gccagagacc ttctggatcg
     2041 gttgtcgagt aaggatctcc aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     2101 cctttcgttt tatctgttgt ttgtcggtga acgctctcta ctagagtcac actggctcac
     2161 cttcgggtgg gcctttctgc gtttatagga tcctaactcg agcctaggga tatattccgc
     2221 ttcctcgctc actgactcgc tacgctcggt cgttcgactg cggcgagcgg aaatggctta
     2281 cgaacggggc ggagatttcc tggaagatgc caggaagata cttaacaggg aagtgagagg
     2341 gccgcggcaa agccgttttt ccataggctc cgcccccctg acaagcatca cgaaatctga
     2401 cgctcaaatc agtggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     2461 ggcggctccc tcgtgcgctc tcctgttcct gcctttcggt ttaccggtgt cattccgctg
     2521 ttatggccgc gtttgtctca ttccacgcct gacactcagt tccgggtagg cagttcgctc
     2581 caagctggac tgtatgcacg aaccccccgt tcagtccgac cgctgcgcct tatccggtaa
     2641 ctatcgtctt gagtccaacc cggaaagaca tgcaaaagca ccactggcag cagccactgg
     2701 taattgattt agaggagtta gtcttgaagt catgcgccgg ttaaggctaa actgaaagga
     2761 caagttttgg tga
//
