LOCUS       sequence-444679-        2747 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and p15A origin of 
            replication. Carries F-G 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_FG
FEATURES             Location/Qualifiers
     source          1..2747
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(26..43)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(160..705)
                     /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(240..259)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(819..913)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(946..1761)
                     /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(946..1761)
                     /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     1669..1688
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     1753..1772
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     1966..1984
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(1988..2031)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    2034..2055
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(2050..2373)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TSL"
     primer_bind     2268..2290
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2282..2299
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2283..2299
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    2300..2356
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(2369..2385)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2369..2385)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2382..2404)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2393..2409
                     /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(2417..2423,2424..2441,2442..2447))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2462..2483
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     terminator      2616..2687
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2703..2730
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
ORIGIN
        1 cgagcctagg gatatattcc gcttcctcgc tcactgactc gctacgctcg gtcgttcgac
       61 tgcggcgagc ggaaatggct tacgaacggg gcggagattt cctggaagat gccaggaaga
      121 tacttaacag ggaagtgaga gggccgcggc aaagccgttt ttccataggc tccgcccccc
      181 tgacaagcat cacgaaatct gacgctcaaa tcagtggtgg cgaaacccga caggactata
      241 aagataccag gcgtttcccc ctggcggctc cctcgtgcgc tctcctgttc ctgcctttcg
      301 gtttaccggt gtcattccgc tgttatggcc gcgtttgtct cattccacgc ctgacactca
      361 gttccgggta ggcagttcgc tccaagctgg actgtatgca cgaacccccc gttcagtccg
      421 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggaaaga catgcaaaag
      481 caccactggc agcagccact ggtaattgat ttagaggagt tagtcttgaa gtcatgcgcc
      541 ggttaaggct aaactgaaag gacaagtttt ggtgactgcg ctcctccaag ccagttacct
      601 cggttcaaag agttggtagc tcagagaacc ttcgaaaaac cgccctgcaa ggcggttttt
      661 tcgttttcag agcaagagat tacgcgcaga ccaaaacgat ctcaagaaga tcatcttatt
      721 aatcagataa aatatttcta gatttcagtg caatttatct cttcaaatgt agcacctgaa
      781 gtcagcccca tacgatataa gttgttacta gtgcttggat tctcaccaat aaaaaacgcc
      841 cggcggcaac cgagcgttct gaacaaatcc agatggagtt ctgaggtcat tactggatct
      901 atcaacagga gtccaagcga gctctcgaac cccagagtcc cgcgattaga aaaactcatc
      961 gagcatcaaa tgaaactgca atttattcat atcaggatta tcaataccat atttttgaaa
     1021 aagccgtttc tgtaatgaag gagaaaactc accgaggcag ttccatagga tggcaagatc
     1081 ctggtatcgg tctgcgattc cgactcgtcc aacatcaata caacctatta atttcccctc
     1141 gtcaaaaata aggttatcaa gtgagaaatc accatgagtg acgactgaat ccggtgagaa
     1201 tggcaaaagc ttatgcattt ctttccagac ttgttcaaca ggccagccat tacgctcgtc
     1261 atcaaaatca ctcgcatcaa ccaaaccgtt attcattcgt gattgcgcct gagcaagacg
     1321 aaatacgcga tcgctgttaa aaggacaatt acaaacagga atcgaatgca accggcgcag
     1381 gaacactgcc agcgcatcaa caatattttc acctgaatca ggatattctt ctaatacctg
     1441 gaatgctgtt ttcccgggga tcgcagtggt gagtaaccat gcatcatcag gagtacggat
     1501 aaaatgcttg atggtcggaa gaggcataaa ttccgtcagc cagtttagtc tgaccatctc
     1561 atctgtaaca tcattggcaa cgctaccttt gccatgtttc agaaacaact ctggcgcatc
     1621 gggcttccca tacaatcgat agattgtcgc acctgattgc ccgacattat cgcgagccca
     1681 tttataccca tataaatcag catccatgtt ggaatttaat cgcggcctgg agcaagacgt
     1741 ttcccgttga atatggctca taacacccct tgtattactg tttatgtaag cagacagttt
     1801 tattgttcat gatgatatat ttttatcttg tgcaatgtaa catcagagat tttgagacac
     1861 aacgtggctt tgttgaataa atcgaacttt tgctgagttg aaggatcagc tcgagtgcca
     1921 cctgacgtct aagaaaccat tattatcatg acattaacct ataaaaatag gcgtatcacg
     1981 aggcagaatt tcagataaaa aaaatcctta gctttcgcta aggatgattt ctggaattcg
     2041 cggccgcttc tagagactag tggaagacat cgctagagac ctgcaccata tgcggtgtga
     2101 aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cgccattcgc cattcaggct
     2161 gcgcaactgt tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa
     2221 agggggatgt gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg
     2281 ttgtaaaacg acggccagtg aattcgagct cggtacccgg ggatcctcta gagtcgacct
     2341 gcaggcatgc aagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     2401 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     2461 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     2521 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc gggggtctct tgccatgtct
     2581 tctggatcgg ttgtcgagta aggatctcca ggcatcaaat aaaacgaaag gctcagtcga
     2641 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctctac tagagtcaca
     2701 ctggctcacc ttcgggtggg cctttctgcg tttataggat cctaact
//
