LOCUS       sequence-468587-        2773 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with ampicillin resistance and p15A 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 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_AH
FEATURES             Location/Qualifiers
     source          1..2773
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(67..161)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(194..985,986..1054))
                     /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"
     primer_bind     817..836
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1055..1159)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     1227..1245
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1249..1292)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1295..1316
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1330..1638)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCREDIS"
     primer_bind     1533..1555
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1547..1564
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1548..1564
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1565..1621
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1634..1650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1634..1650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1647..1669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1658..1674
                     /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(1682..1688,1689..1706,1707..1712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1727..1748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1890..1961
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1977..2004
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(2047..2064)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2181..2726)
                     /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(2261..2280)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 atttatctct tcaaatgtag cacctgaagt cagccccata cgatataagt tgttactagt
       61 gcttggattc tcaccaataa aaaacgcccg gcggcaaccg agcgttctga acaaatccag
      121 atggagttct gaggtcatta ctggatctat caacaggagt ccaagcgagc tctcgaaccc
      181 cagagtcccg cgattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
      241 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
      301 taccatctgg ccccagtgct gcaatgatac cgcgcgaccc acgctcaccg gctccagatt
      361 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
      421 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
      481 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
      541 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
      601 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
      661 cagtgttatc actaatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
      721 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
      781 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
      841 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
      901 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
      961 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     1021 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     1081 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     1141 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     1201 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat ttcagataaa
     1261 aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt ctagagacta
     1321 gtggaggtct caggagatgt cttccctgca ccatatgcgg tgtgaaatac cgcacagatg
     1381 cgtaaggaga aaataccgca tcaggcgcca ttcgccattc aggctgcgca actgttggga
     1441 agggcgatcg gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc
     1501 aaggcgatta agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc
     1561 cagtgaattc gagctcggta cccggggatc ctctagagtc gacctgcagg catgcaagct
     1621 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     1681 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     1741 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     1801 tgcattaatg aatcggccaa cgcgcggggg ttaagaagac gtactaagag accttctgga
     1861 tcggttgtcg agtaaggatc tccaggcatc aaataaaacg aaaggctcag tcgaaagact
     1921 gggcctttcg ttttatctgt tgtttgtcgg tgaacgctct ctactagagt cacactggct
     1981 caccttcggg tgggcctttc tgcgtttata ggatcctaac tcgagcctag ggatatattc
     2041 cgcttcctcg ctcactgact cgctacgctc ggtcgttcga ctgcggcgag cggaaatggc
     2101 ttacgaacgg ggcggagatt tcctggaaga tgccaggaag atacttaaca gggaagtgag
     2161 agggccgcgg caaagccgtt tttccatagg ctccgccccc ctgacaagca tcacgaaatc
     2221 tgacgctcaa atcagtggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2281 cctggcggct ccctcgtgcg ctctcctgtt cctgcctttc ggtttaccgg tgtcattccg
     2341 ctgttatggc cgcgtttgtc tcattccacg cctgacactc agttccgggt aggcagttcg
     2401 ctccaagctg gactgtatgc acgaaccccc cgttcagtcc gaccgctgcg ccttatccgg
     2461 taactatcgt cttgagtcca acccggaaag acatgcaaaa gcaccactgg cagcagccac
     2521 tggtaattga tttagaggag ttagtcttga agtcatgcgc cggttaaggc taaactgaaa
     2581 ggacaagttt tggtgactgc gctcctccaa gccagttacc tcggttcaaa gagttggtag
     2641 ctcagagaac cttcgaaaaa ccgccctgca aggcggtttt ttcgttttca gagcaagaga
     2701 ttacgcgcag accaaaacga tctcaagaag atcatcttat taatcagata aaatatttct
     2761 agatttcagt gca
//
