LOCUS       sequence-442574-        3156 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Destination vector with ampicillin resistance and pBR322 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 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 Apr 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_AG
FEATURES             Location/Qualifiers
     source          1..3156
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     476..495
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(714..818)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     886..904
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(908..951)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    954..975
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(989..1297)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1192..1214
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1206..1223
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1207..1223
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1224..1280
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1293..1309)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1293..1309)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1306..1328)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1317..1333
                     /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(1341..1347,1348..1365,1366..1371))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1386..1407
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1727..1918
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1912..1934)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    2020..2159
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2075..2094
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2174..2191)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2345..2933)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2425..2444)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2964..3003)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(3009..3156,1..644,645..713))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITISDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
       61 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
      121 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
      181 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
      241 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
      301 gtcagaagta agttggccgc agtgttatca ctaatggtta tggcagcact gcataattct
      361 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
      421 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
      481 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
      541 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
      601 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
      661 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
      721 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
      781 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
      841 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
      901 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
      961 cggccgcttc tagagactag tggaggtctc aggagatgtc ttccctgcac catatgcggt
     1021 gtgaaatacc gcacagatgc gtaaggagaa aataccgcat caggcgccat tcgccattca
     1081 ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg
     1141 cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt tcccagtcac
     1201 gacgttgtaa aacgacggcc agtgaattcg agctcggtac ccggggatcc tctagagtcg
     1261 acctgcaggc atgcaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt
     1321 tatccgctca caattccaca caacatacga gccggaagca taaagtgtaa agcctggggt
     1381 gcctaatgag tgagctaact cacattaatt gcgttgcgct cactgcccgc tttccagtcg
     1441 ggaaacctgt cgtgccagct gcattaatga atcggccaac gcgcgggggt taagaagacg
     1501 ttgccagaga ccttctaaca cctacatctg tattaacgaa gcgctggcat tgaccctgag
     1561 tgatttttct ctggtcccgc cgcatccata ccgccagttg tttaccctca caacgttcca
     1621 gtaaccgggc atgttcatca tcagtaaccc gtatcgtgag catcctctct cgtttcatcg
     1681 gtatcattac cccatgaaca gaaatccccc ttacacggag gcatcagtga ccaaacagga
     1741 aaaaaccgcc cttaacatgg cccgctttat cagaagccag acattaacgc ttctggagaa
     1801 actcaacgag ctggacgcgg atgaacaggc agacatctgt gaatcgcttc acgaccacgc
     1861 tgatgagctt taccgcagct gcctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     1921 catgcagctc ccggcgacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     1981 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc gcagccatga cccagtcacg
     2041 tagcgatagc ggagtgtata ctggcttaac tatgcggcat cagagcagat tgtactgaga
     2101 gtgcaccatt gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc
     2161 gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg
     2221 tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa
     2281 agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg
     2341 cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga
     2401 ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg
     2461 tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg
     2521 gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc
     2581 gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg
     2641 gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca
     2701 ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt
     2761 ggcctaacta cggctacact agaaggacag tatttggtat ctgcgctctg ctgaagccag
     2821 ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg
     2881 gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc
     2941 ctttgatctt ttctaccggt ccgtccaaaa aaaaaggctc caaaaggagc ctttaattgt
     3001 atcggtgatt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3061 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3121 tctggcccca gtgctgcaat gataccgcgc gaccca
//
