LOCUS       sequence-442595-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and p15A origin of 
            replication. Carries A-F 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_AF
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(48..64)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(48..64)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(61..83)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    72..88
                     /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(96..102,103..120,121..126))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    141..162
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      295..366
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      382..409
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(452..469)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(586..1131)
                     /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(666..685)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1245..1339)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1372..2187)
                     /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(1372..2187)
                     /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"
     primer_bind     2095..2114
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2179..2198
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2392..2410
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2414..2457)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2460..2481
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(2476..2747,1..52))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TSL"
     primer_bind     2694..2716
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2708..2725
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2709..2725
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(2726..2747,1..35)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 gatcctctag agtcgacctg caggcatgca agcttggcgt aatcatggtc atagctgttt
       61 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
      121 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
      181 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
      241 ggggtctctc gctatgtctt ctggatcggt tgtcgagtaa ggatctccag gcatcaaata
      301 aaacgaaagg ctcagtcgaa agactgggcc tttcgtttta tctgttgttt gtcggtgaac
      361 gctctctact agagtcacac tggctcacct tcgggtgggc ctttctgcgt ttataggatc
      421 ctaactcgag cctagggata tattccgctt cctcgctcac tgactcgcta cgctcggtcg
      481 ttcgactgcg gcgagcggaa atggcttacg aacggggcgg agatttcctg gaagatgcca
      541 ggaagatact taacagggaa gtgagagggc cgcggcaaag ccgtttttcc ataggctccg
      601 cccccctgac aagcatcacg aaatctgacg ctcaaatcag tggtggcgaa acccgacagg
      661 actataaaga taccaggcgt ttccccctgg cggctccctc gtgcgctctc ctgttcctgc
      721 ctttcggttt accggtgtca ttccgctgtt atggccgcgt ttgtctcatt ccacgcctga
      781 cactcagttc cgggtaggca gttcgctcca agctggactg tatgcacgaa ccccccgttc
      841 agtccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gaaagacatg
      901 caaaagcacc actggcagca gccactggta attgatttag aggagttagt cttgaagtca
      961 tgcgccggtt aaggctaaac tgaaaggaca agttttggtg actgcgctcc tccaagccag
     1021 ttacctcggt tcaaagagtt ggtagctcag agaaccttcg aaaaaccgcc ctgcaaggcg
     1081 gttttttcgt tttcagagca agagattacg cgcagaccaa aacgatctca agaagatcat
     1141 cttattaatc agataaaata tttctagatt tcagtgcaat ttatctcttc aaatgtagca
     1201 cctgaagtca gccccatacg atataagttg ttactagtgc ttggattctc accaataaaa
     1261 aacgcccggc ggcaaccgag cgttctgaac aaatccagat ggagttctga ggtcattact
     1321 ggatctatca acaggagtcc aagcgagctc tcgaacccca gagtcccgcg attagaaaaa
     1381 ctcatcgagc atcaaatgaa actgcaattt attcatatca ggattatcaa taccatattt
     1441 ttgaaaaagc cgtttctgta atgaaggaga aaactcaccg aggcagttcc ataggatggc
     1501 aagatcctgg tatcggtctg cgattccgac tcgtccaaca tcaatacaac ctattaattt
     1561 cccctcgtca aaaataaggt tatcaagtga gaaatcacca tgagtgacga ctgaatccgg
     1621 tgagaatggc aaaagcttat gcatttcttt ccagacttgt tcaacaggcc agccattacg
     1681 ctcgtcatca aaatcactcg catcaaccaa accgttattc attcgtgatt gcgcctgagc
     1741 aagacgaaat acgcgatcgc tgttaaaagg acaattacaa acaggaatcg aatgcaaccg
     1801 gcgcaggaac actgccagcg catcaacaat attttcacct gaatcaggat attcttctaa
     1861 tacctggaat gctgttttcc cggggatcgc agtggtgagt aaccatgcat catcaggagt
     1921 acggataaaa tgcttgatgg tcggaagagg cataaattcc gtcagccagt ttagtctgac
     1981 catctcatct gtaacatcat tggcaacgct acctttgcca tgtttcagaa acaactctgg
     2041 cgcatcgggc ttcccataca atcgatagat tgtcgcacct gattgcccga cattatcgcg
     2101 agcccattta tacccatata aatcagcatc catgttggaa tttaatcgcg gcctggagca
     2161 agacgtttcc cgttgaatat ggctcataac accccttgta ttactgttta tgtaagcaga
     2221 cagttttatt gttcatgatg atatattttt atcttgtgca atgtaacatc agagattttg
     2281 agacacaacg tggctttgtt gaataaatcg aacttttgct gagttgaagg atcagctcga
     2341 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     2401 atcacgaggc agaatttcag ataaaaaaaa tccttagctt tcgctaagga tgatttctgg
     2461 aattcgcggc cgcttctaga gactagtgga agacatggag agagacctgc accatatgcg
     2521 gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc attcgccatt
     2581 caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct
     2641 ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc
     2701 acgacgttgt aaaacgacgg ccagtgaatt cgagctcggt acccggg
//
