LOCUS       sequence-442035-        3156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and pBR322 origin of 
            replication. Carries A-F 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 3156)
  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 3156)
  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_pBR322_AF
FEATURES             Location/Qualifiers
     source          1..3156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(262..301)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(307..1098,1099..1167))
                     /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     930..949
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1168..1272)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1340..1358
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1362..1405)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1408..1429
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1443..1751)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1646..1668
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1660..1677
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1661..1677
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1678..1734
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1747..1763)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1747..1763)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1760..1782)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1771..1787
                     /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(1795..1801,1802..1819,1820..1825))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1840..1861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             2181..2372
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(2366..2388)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2474..2613
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2529..2548
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2628..2645)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(join(2799..3156,1..231))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2879..2898)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
       61 cctaactacg gctacactag aaggacagta tttggtatct gcgctctgct gaagccagtt
      121 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
      181 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
      241 ttgatctttt ctaccggtcc gtccaaaaaa aaaggctcca aaaggagcct ttaattgtat
      301 cggtgattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
      361 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
      421 tggccccagt gctgcaatga taccgcgcga cccacgctca ccggctccag atttatcagc
      481 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
      541 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
      601 gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
      661 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
      721 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
      781 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
      841 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
      901 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
      961 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     1021 tagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     1081 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     1141 ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta
     1201 tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa ataaacaaat
     1261 aggggttccg cgcacatttc cccgaaaagt gccacctgac gtctaagaaa ccattattat
     1321 catgacatta acctataaaa ataggcgtat cacgaggcag aatttcagat aaaaaaaatc
     1381 cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagaga ctagtggagg
     1441 tctcaggaga tgtcttccct gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     1501 agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc gcaactgttg ggaagggcga
     1561 tcggtgcggg cctcttcgct attacgccag ctggcgaaag ggggatgtgc tgcaaggcga
     1621 ttaagttggg taacgccagg gttttcccag tcacgacgtt gtaaaacgac ggccagtgaa
     1681 ttcgagctcg gtacccgggg atcctctaga gtcgacctgc aggcatgcaa gcttggcgta
     1741 atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat
     1801 acgagccgga agcataaagt gtaaagcctg gggtgcctaa tgagtgagct aactcacatt
     1861 aattgcgttg cgctcactgc ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta
     1921 atgaatcggc caacgcgcgg gggttaagaa gacgtcgcta gagaccttct aacacctaca
     1981 tctgtattaa cgaagcgctg gcattgaccc tgagtgattt ttctctggtc ccgccgcatc
     2041 cataccgcca gttgtttacc ctcacaacgt tccagtaacc gggcatgttc atcatcagta
     2101 acccgtatcg tgagcatcct ctctcgtttc atcggtatca ttaccccatg aacagaaatc
     2161 ccccttacac ggaggcatca gtgaccaaac aggaaaaaac cgcccttaac atggcccgct
     2221 ttatcagaag ccagacatta acgcttctgg agaaactcaa cgagctggac gcggatgaac
     2281 aggcagacat ctgtgaatcg cttcacgacc acgctgatga gctttaccgc agctgcctcg
     2341 cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggcg acggtcacag
     2401 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     2461 gcgggtgtcg gggcgcagcc atgacccagt cacgtagcga tagcggagtg tatactggct
     2521 taactatgcg gcatcagagc agattgtact gagagtgcac cattgcggtg tgaaataccg
     2581 cacagatgcg taaggagaaa ataccgcatc aggcgctctt ccgcttcctc gctcactgac
     2641 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     2701 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     2761 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     2821 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     2881 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     2941 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     3001 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     3061 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     3121 gtaagacacg acttatcgcc actggcagca gccact
//
