LOCUS       sequence-442584-        2773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and p15A 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 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_EH
FEATURES             Location/Qualifiers
     source          1..2773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     258..277
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(496..600)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     668..686
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(690..733)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    736..757
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(771..1079)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     974..996
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     988..1005
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     989..1005
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1006..1062
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1075..1091)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1075..1091)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1088..1110)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1099..1115
                     /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(1123..1129,1130..1147,1148..1153))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1168..1189
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1331..1402
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1418..1445
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1488..1505)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1622..2167)
                     /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(1702..1721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2281..2375)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2408..2773,1..426,427..495))
                     /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 cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag
       61 ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt
      121 tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac
      181 tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg
      241 cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat
      301 tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc
      361 gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc
      421 tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa
      481 atgttgaata ctcatactct tcctttttca atattattga agcatttatc agggttattg
      541 tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg
      601 cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac
      661 ctataaaaat aggcgtatca cgaggcagaa tttcagataa aaaaaatcct tagctttcgc
      721 taaggatgat ttctggaatt cgcggccgct tctagagact agtggaggtc tcagcttatg
      781 tcttccctgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
      841 atcaggcgcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
      901 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
      961 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt
     1021 acccggggat cctctagagt cgacctgcag gcatgcaagc ttggcgtaat catggtcata
     1081 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     1141 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     1201 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     1261 acgcgcgggg gttaagaaga cgtactaaga gaccttctgg atcggttgtc gagtaaggat
     1321 ctccaggcat caaataaaac gaaaggctca gtcgaaagac tgggcctttc gttttatctg
     1381 ttgtttgtcg gtgaacgctc tctactagag tcacactggc tcaccttcgg gtgggccttt
     1441 ctgcgtttat aggatcctaa ctcgagccta gggatatatt ccgcttcctc gctcactgac
     1501 tcgctacgct cggtcgttcg actgcggcga gcggaaatgg cttacgaacg gggcggagat
     1561 ttcctggaag atgccaggaa gatacttaac agggaagtga gagggccgcg gcaaagccgt
     1621 ttttccatag gctccgcccc cctgacaagc atcacgaaat ctgacgctca aatcagtggt
     1681 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggcggc tccctcgtgc
     1741 gctctcctgt tcctgccttt cggtttaccg gtgtcattcc gctgttatgg ccgcgtttgt
     1801 ctcattccac gcctgacact cagttccggg taggcagttc gctccaagct ggactgtatg
     1861 cacgaacccc ccgttcagtc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     1921 aacccggaaa gacatgcaaa agcaccactg gcagcagcca ctggtaattg atttagagga
     1981 gttagtcttg aagtcatgcg ccggttaagg ctaaactgaa aggacaagtt ttggtgactg
     2041 cgctcctcca agccagttac ctcggttcaa agagttggta gctcagagaa ccttcgaaaa
     2101 accgccctgc aaggcggttt tttcgttttc agagcaagag attacgcgca gaccaaaacg
     2161 atctcaagaa gatcatctta ttaatcagat aaaatatttc tagatttcag tgcaatttat
     2221 ctcttcaaat gtagcacctg aagtcagccc catacgatat aagttgttac tagtgcttgg
     2281 attctcacca ataaaaaacg cccggcggca accgagcgtt ctgaacaaat ccagatggag
     2341 ttctgaggtc attactggat ctatcaacag gagtccaagc gagctctcga accccagagt
     2401 cccgcgatta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     2461 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     2521 ctggccccag tgctgcaatg ataccgcgcg acccacgctc accggctcca gatttatcag
     2581 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     2641 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     2701 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     2761 cttcattcag ctc
//
