LOCUS       sequence-442592-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and p15A origin of 
            replication. Carries A-E CIDAR MoClo overhangs for level 1 construct
            cloning..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 2747)
  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 2747)
  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: DVK_p15A_AE
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     467..486
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     551..570
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     764..782
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(786..829)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    832..853
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(848..1171)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TSL"
     primer_bind     1066..1088
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1080..1097
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1081..1097
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1098..1154
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1167..1183)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1167..1183)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1180..1202)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1191..1207
                     /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(1215..1221,1222..1239,1240..1245))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1260..1281
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1414..1485
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1501..1528
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1571..1588)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1705..2250)
                     /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(1785..1804)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2364..2458)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(join(2491..2747,1..559))
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     CDS             complement(join(2491..2747,1..559))
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
ORIGIN
        1 gcaaaagctt atgcatttct ttccagactt gttcaacagg ccagccatta cgctcgtcat
       61 caaaatcact cgcatcaacc aaaccgttat tcattcgtga ttgcgcctga gcaagacgaa
      121 atacgcgatc gctgttaaaa ggacaattac aaacaggaat cgaatgcaac cggcgcagga
      181 acactgccag cgcatcaaca atattttcac ctgaatcagg atattcttct aatacctgga
      241 atgctgtttt cccggggatc gcagtggtga gtaaccatgc atcatcagga gtacggataa
      301 aatgcttgat ggtcggaaga ggcataaatt ccgtcagcca gtttagtctg accatctcat
      361 ctgtaacatc attggcaacg ctacctttgc catgtttcag aaacaactct ggcgcatcgg
      421 gcttcccata caatcgatag attgtcgcac ctgattgccc gacattatcg cgagcccatt
      481 tatacccata taaatcagca tccatgttgg aatttaatcg cggcctggag caagacgttt
      541 cccgttgaat atggctcata acaccccttg tattactgtt tatgtaagca gacagtttta
      601 ttgttcatga tgatatattt ttatcttgtg caatgtaaca tcagagattt tgagacacaa
      661 cgtggctttg ttgaataaat cgaacttttg ctgagttgaa ggatcagctc gagtgccacc
      721 tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag
      781 gcagaatttc agataaaaaa aatccttagc tttcgctaag gatgatttct ggaattcgcg
      841 gccgcttcta gagactagtg gaagacatgg agagagacct gcaccatatg cggtgtgaaa
      901 taccgcacag atgcgtaagg agaaaatacc gcatcaggcg ccattcgcca ttcaggctgc
      961 gcaactgttg ggaagggcga tcggtgcggg cctcttcgct attacgccag ctggcgaaag
     1021 ggggatgtgc tgcaaggcga ttaagttggg taacgccagg gttttcccag tcacgacgtt
     1081 gtaaaacgac ggccagtgaa ttcgagctcg gtacccgggg atcctctaga gtcgacctgc
     1141 aggcatgcaa gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa ttgttatccg
     1201 ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg gggtgcctaa
     1261 tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca gtcgggaaac
     1321 ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg gggtctctgc ttatgtcttc
     1381 tggatcggtt gtcgagtaag gatctccagg catcaaataa aacgaaaggc tcagtcgaaa
     1441 gactgggcct ttcgttttat ctgttgtttg tcggtgaacg ctctctacta gagtcacact
     1501 ggctcacctt cgggtgggcc tttctgcgtt tataggatcc taactcgagc ctagggatat
     1561 attccgcttc ctcgctcact gactcgctac gctcggtcgt tcgactgcgg cgagcggaaa
     1621 tggcttacga acggggcgga gatttcctgg aagatgccag gaagatactt aacagggaag
     1681 tgagagggcc gcggcaaagc cgtttttcca taggctccgc ccccctgaca agcatcacga
     1741 aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga ctataaagat accaggcgtt
     1801 tccccctggc ggctccctcg tgcgctctcc tgttcctgcc tttcggttta ccggtgtcat
     1861 tccgctgtta tggccgcgtt tgtctcattc cacgcctgac actcagttcc gggtaggcag
     1921 ttcgctccaa gctggactgt atgcacgaac cccccgttca gtccgaccgc tgcgccttat
     1981 ccggtaacta tcgtcttgag tccaacccgg aaagacatgc aaaagcacca ctggcagcag
     2041 ccactggtaa ttgatttaga ggagttagtc ttgaagtcat gcgccggtta aggctaaact
     2101 gaaaggacaa gttttggtga ctgcgctcct ccaagccagt tacctcggtt caaagagttg
     2161 gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg ttttttcgtt ttcagagcaa
     2221 gagattacgc gcagaccaaa acgatctcaa gaagatcatc ttattaatca gataaaatat
     2281 ttctagattt cagtgcaatt tatctcttca aatgtagcac ctgaagtcag ccccatacga
     2341 tataagttgt tactagtgct tggattctca ccaataaaaa acgcccggcg gcaaccgagc
     2401 gttctgaaca aatccagatg gagttctgag gtcattactg gatctatcaa caggagtcca
     2461 agcgagctct cgaaccccag agtcccgcga ttagaaaaac tcatcgagca tcaaatgaaa
     2521 ctgcaattta ttcatatcag gattatcaat accatatttt tgaaaaagcc gtttctgtaa
     2581 tgaaggagaa aactcaccga ggcagttcca taggatggca agatcctggt atcggtctgc
     2641 gattccgact cgtccaacat caatacaacc tattaatttc ccctcgtcaa aaataaggtt
     2701 atcaagtgag aaatcaccat gagtgacgac tgaatccggt gagaatg
//
