LOCUS       sequence-444824-        3157 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and pBR322 origin of 
            replication. Carries A-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 3157)
  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 3157)
  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_AH
FEATURES             Location/Qualifiers
     source          1..3157
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     445..464
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(683..787)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     855..873
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(877..920)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    923..944
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(958..1266)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1161..1183
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1175..1192
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1176..1192
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1193..1249
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1262..1278)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1262..1278)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1275..1297)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1286..1302
                     /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(1310..1316,1317..1334,1335..1340))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1355..1376
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1882..1904)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1990..2129
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     2045..2064
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2144..2161)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2315..2903)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2395..2414)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2934..2973)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2979..3157,1..613,614..682))
                     /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 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
       61 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
      121 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
      181 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
      241 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
      301 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
      361 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
      421 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
      481 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
      541 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
      601 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
      661 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
      721 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
      781 ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt attatcatga
      841 cattaaccta taaaaatagg cgtatcacga ggcagaattt cagataaaaa aaatccttag
      901 ctttcgctaa ggatgatttc tggaattcgc ggccgcttct agagactagt ggaggtctca
      961 ggagatgtct tccctgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     1021 ataccgcatc aggcgccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt
     1081 gcgggcctct tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag
     1141 ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaattcga
     1201 gctcggtacc cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat
     1261 ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag
     1321 ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg
     1381 cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa
     1441 tcggccaacg cgcgggggtt aagaagacgt actaagagac cttctaacac ctacatctgt
     1501 attaacgaag cgctggcatt gaccctgagt gatttttctc tggtcccgcc gcatccatac
     1561 cgccagttgt ttaccctcac aacgttccag taaccgggca tgttcatcat cagtaacccg
     1621 tatcgtgagc atcctctctc gtttcatcgg tatcattacc ccatgaacag aaatccccct
     1681 tacacggagg catcagtgac caaacaggaa aaaaccgccc ttaacatggc ccgctttatc
     1741 agaagccaga cattaacgct tctggagaaa ctcaacgagc tggaccgcgg atgaacaggc
     1801 agacatctgt gaatcgcttc acgaccacgc tgatgagctt taccgcagct gcctcgcgcg
     1861 tttcggtgat gacggtgaaa acctctgaca catgcagctc ccggcgacgg tcacagcttg
     1921 tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg
     1981 gtgtcggggc gcagccatga cccagtcacg tagcgatagc ggagtgtata ctggcttaac
     2041 tatgcggcat cagagcagat tgtactgaga gtgcaccatt gcggtgtgaa ataccgcaca
     2101 gatgcgtaag gagaaaatac cgcatcaggc gctcttccgc ttcctcgctc actgactcgc
     2161 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     2221 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     2281 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     2341 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     2401 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     2461 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     2521 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     2581 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     2641 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     2701 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaaggacag
     2761 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     2821 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     2881 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctaccggt ccgtccaaaa
     2941 aaaaaggctc caaaaggagc ctttaattgt atcggtgatt accaatgctt aatcagtgag
     3001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     3061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcgc
     3121 gacccacgct caccggctcc agatttatca gcaataa
//
