LOCUS       sequence-442590-        2773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and p15A 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 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_AF
FEATURES             Location/Qualifiers
     source          1..2773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(109..900,901..969))
                     /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     732..751
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(970..1074)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1142..1160
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1164..1207)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1210..1231
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1245..1553)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1448..1470
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1462..1479
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1463..1479
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1480..1536
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1549..1565)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1549..1565)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1562..1584)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1573..1589
                     /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(1597..1603,1604..1621,1622..1627))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1642..1663
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1805..1876
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1892..1919
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1962..1979)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2096..2641)
                     /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(2176..2195)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(join(2755..2773,1..76))
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 gcccggcggc aaccgagcgt tctgaacaaa tccagatgga gttctgaggt cattactgga
       61 tctatcaaca ggagtccaag cgagctctcg aaccccagag tcccgcgatt accaatgctt
      121 aatcagtgag gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact
      181 ccccgtcgtg tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat
      241 gataccgcgc gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg
      301 aagggccgag cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg
      361 ttgccgggaa gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat
      421 tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc
      481 ccaacgatca aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt
      541 cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc
      601 agcactgcat aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga
      661 gtactcaacc aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc
      721 gtcaatacgg gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa
      781 acgttcttcg gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta
      841 acccactcgt gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg
      901 agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg
      961 aatactcata ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat
     1021 gagcggatac atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt
     1081 tccccgaaaa gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa
     1141 aaataggcgt atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga
     1201 tgatttctgg aattcgcggc cgcttctaga gactagtgga ggtctcagga gatgtcttcc
     1261 ctgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1321 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1381 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1441 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     1501 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     1561 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1621 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1681 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1741 gggggttaag aagacgtcgc tagagacctt ctggatcggt tgtcgagtaa ggatctccag
     1801 gcatcaaata aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt
     1861 gtcggtgaac gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt
     1921 ttataggatc ctaactcgag cctagggata tattccgctt cctcgctcac tgactcgcta
     1981 cgctcggtcg ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg
     2041 gaagatgcca ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc
     2101 ataggctccg cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa
     2161 acccgacagg actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc
     2221 ctgttcctgc ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt
     2281 ccacgcctga cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa
     2341 ccccccgttc agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     2401 gaaagacatg caaaagcacc actggcagca gccactggta attgatttag aggagttagt
     2461 cttgaagtca tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc
     2521 tccaagccag ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc
     2581 ctgcaaggcg gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca
     2641 agaagatcat cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc
     2701 aaatgtagca cctgaagtca gccccatacg atataagttg ttactagtgc ttggattctc
     2761 accaataaaa aac
//
