LOCUS       sequence-442588-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and p15A origin of 
            replication. Carries G-H 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_GH
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(52..68)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(52..68)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(65..87)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    76..92
                     /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(100..106,107..124,125..130))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    145..166
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      299..370
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      386..413
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     primer_bind     complement(456..473)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(590..1135)
                     /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(670..689)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1249..1343)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(1376..2191)
                     /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(1376..2191)
                     /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     2099..2118
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     2183..2202
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     2396..2414
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(2418..2461)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2464..2485
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(join(2480..2747,1..56))
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TSL"
     primer_bind     2698..2720
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2712..2729
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2713..2729
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    join(2730..2747,1..39)
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
ORIGIN
        1 cggggatcct ctagagtcga cctgcaggca tgcaagcttg gcgtaatcat ggtcatagct
       61 gtttcctgtg tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat
      121 aaagtgtaaa gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc
      181 actgcccgct ttccagtcgg gaaacctgtc gtgccagctg cattaatgaa tcggccaacg
      241 cgcgggggtc tctactaatg tcttctggat cggttgtcga gtaaggatct ccaggcatca
      301 aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt gtttgtcggt
      361 gaacgctctc tactagagtc acactggctc accttcgggt gggcctttct gcgtttatag
      421 gatcctaact cgagcctagg gatatattcc gcttcctcgc tcactgactc gctacgctcg
      481 gtcgttcgac tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat
      541 gccaggaaga tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc
      601 tccgcccccc tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga
      661 caggactata aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc
      721 ctgcctttcg gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc
      781 ctgacactca gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc
      841 gttcagtccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga
      901 catgcaaaag caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa
      961 gtcatgcgcc ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag
     1021 ccagttacct cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa
     1081 ggcggttttt tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga
     1141 tcatcttatt aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt
     1201 agcacctgaa gtcagcccca tacgatataa gttgttacta gtgcttggat tctcaccaat
     1261 aaaaaacgcc cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat
     1321 tactggatct atcaacagga gtccaagcga gctctcgaac cccagagtcc cgcgattaga
     1381 aaaactcatc gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat
     1441 atttttgaaa aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga
     1501 tggcaagatc ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta
     1561 atttcccctc gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat
     1621 ccggtgagaa tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat
     1681 tacgctcgtc atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct
     1741 gagcaagacg aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca
     1801 accggcgcag gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt
     1861 ctaatacctg gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag
     1921 gagtacggat aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc
     1981 tgaccatctc atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact
     2041 ctggcgcatc gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat
     2101 cgcgagccca tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg
     2161 agcaagacgt ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag
     2221 cagacagttt tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat
     2281 tttgagacac aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagc
     2341 tcgagtgcca cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag
     2401 gcgtatcacg aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt
     2461 ctggaattcg cggccgcttc tagagactag tggaagacat tgccagagac ctgcaccata
     2521 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc
     2581 cattcaggct gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc
     2641 agctggcgaa agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc
     2701 agtcacgacg ttgtaaaacg acggccagtg aattcgagct cggtacc
//
