LOCUS       sequence-442034-        3156 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and pBR322 origin of 
            replication. Carries E-H 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_EH
FEATURES             Location/Qualifiers
     source          1..3156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     486..505
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(724..828)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     896..914
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(918..961)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    964..985
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(999..1307)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1202..1224
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1216..1233
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1217..1233
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1234..1290
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1303..1319)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1303..1319)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1316..1338)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1327..1343
                     /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(1351..1357,1358..1375,1376..1381))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1396..1417
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1737..1928
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1922..1944)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2030..2169
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2085..2104
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2184..2201)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2355..2943)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2435..2454)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2974..3013)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(3019..3156,1..654,655..723))
                     /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"
ORIGIN
        1 gcgcgaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
       61 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
      121 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
      181 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
      241 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
      301 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
      361 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
      421 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
      481 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
      541 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
      601 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
      661 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
      721 catactcttc ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg
      781 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
      841 aaaagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
      901 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
      961 ctggaattcg cggccgcttc tagagactag tggaggtctc agcttatgtc ttccctgcac
     1021 catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat
     1081 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta
     1141 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt
     1201 tcccagtcac gacgttgtaa aacgacggcc agtgaattcg agctcggtac ccggggatcc
     1261 tctagagtcg acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt
     1321 gtgaaattgt tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa
     1381 agcctggggt gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc
     1441 tttccagtcg ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcgggggt
     1501 taagaagacg tactaagaga ccttctaaca cctacatctg tattaacgaa gcgctggcat
     1561 tgaccctgag tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca
     1621 caacgttcca gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct
     1681 cgtttcatcg gtatcattac cccatgaaca gaaatccccc ttacacggag gcatcagtga
     1741 ccaaacagga aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc
     1801 ttctggagaa actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc
     1861 acgaccacgc tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa
     1921 acctctgaca catgcagctc ccggcgacgg tcacagcttg tctgtaagcg gatgccggga
     1981 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga
     2041 cccagtcacg tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat
     2101 tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     2161 cgcatcaggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     2221 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     2281 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     2341 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     2401 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     2461 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     2521 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     2581 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     2641 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     2701 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     2761 ttgaagtggt ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg
     2821 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     2881 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     2941 caagaagatc ctttgatctt ttctaccggt ccgtccaaaa aaaaaggctc caaaaggagc
     3001 ctttaattgt atcggtgatt accaatgctt aatcagtgag gcacctatct cagcgatctg
     3061 tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga
     3121 gggcttacca tctggcccca gtgctgcaat gatacc
//
