LOCUS       sequence-442587-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and p15A origin of 
            replication. Carries E-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_EF
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(6..821)
                     /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(6..821)
                     /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     729..748
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     813..832
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1026..1044
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1048..1091)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1094..1115
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(1110..1433)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLSMSS
                     TSL"
     primer_bind     1328..1350
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     1342..1359
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1343..1359
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1360..1416
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1429..1445)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1429..1445)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1442..1464)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1453..1469
                     /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(1477..1483,1484..1501,1502..1507))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1522..1543
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      1676..1747
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1763..1790
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(1833..1850)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1967..2512)
                     /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(2047..2066)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2626..2720)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 cgcgattaga aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta
       61 tcaataccat atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag
      121 ttccatagga tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata
      181 caacctatta atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg
      241 acgactgaat ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca
      301 ggccagccat tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt
      361 gattgcgcct gagcaagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga
      421 atcgaatgca accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca
      481 ggatattctt ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat
      541 gcatcatcag gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc
      601 cagtttagtc tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc
      661 agaaacaact ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc
      721 ccgacattat cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat
      781 cgcggcctgg agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg
      841 tttatgtaag cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa
      901 catcagagat tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg
      961 aaggatcagc tcgagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct
     1021 ataaaaatag gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta
     1081 aggatgattt ctggaattcg cggccgcttc tagagactag tggaagacat gcttagagac
     1141 ctgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg
     1201 cgccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg
     1261 ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca
     1321 gggttttccc agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacccgg
     1381 ggatcctcta gagtcgacct gcaggcatgc aagcttggcg taatcatggt catagctgtt
     1441 tcctgtgtga aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa
     1501 gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact
     1561 gcccgctttc cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc
     1621 gggggtctct cgctatgtct tctggatcgg ttgtcgagta aggatctcca ggcatcaaat
     1681 aaaacgaaag gctcagtcga aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa
     1741 cgctctctac tagagtcaca ctggctcacc ttcgggtggg cctttctgcg tttataggat
     1801 cctaactcga gcctagggat atattccgct tcctcgctca ctgactcgct acgctcggtc
     1861 gttcgactgc ggcgagcgga aatggcttac gaacggggcg gagatttcct ggaagatgcc
     1921 aggaagatac ttaacaggga agtgagaggg ccgcggcaaa gccgtttttc cataggctcc
     1981 gcccccctga caagcatcac gaaatctgac gctcaaatca gtggtggcga aacccgacag
     2041 gactataaag ataccaggcg tttccccctg gcggctccct cgtgcgctct cctgttcctg
     2101 cctttcggtt taccggtgtc attccgctgt tatggccgcg tttgtctcat tccacgcctg
     2161 acactcagtt ccgggtaggc agttcgctcc aagctggact gtatgcacga accccccgtt
     2221 cagtccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggaaagacat
     2281 gcaaaagcac cactggcagc agccactggt aattgattta gaggagttag tcttgaagtc
     2341 atgcgccggt taaggctaaa ctgaaaggac aagttttggt gactgcgctc ctccaagcca
     2401 gttacctcgg ttcaaagagt tggtagctca gagaaccttc gaaaaaccgc cctgcaaggc
     2461 ggttttttcg ttttcagagc aagagattac gcgcagacca aaacgatctc aagaagatca
     2521 tcttattaat cagataaaat atttctagat ttcagtgcaa tttatctctt caaatgtagc
     2581 acctgaagtc agccccatac gatataagtt gttactagtg cttggattct caccaataaa
     2641 aaacgcccgg cggcaaccga gcgttctgaa caaatccaga tggagttctg aggtcattac
     2701 tggatctatc aacaggagtc caagcgagct ctcgaacccc agagtcc
//
