LOCUS       sequence-450353-        3130 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Destination vector with kanamycin resistance and pBR322 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 3130)
  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 3130)
  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_pBR322_FG
FEATURES             Location/Qualifiers
     source          1..3130
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     340..359
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     424..443
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     637..655
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      complement(659..702)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    705..726
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     CDS             complement(721..1044)
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITPSLHACRSTLEDPRVPSSNSLAVVLQRRDWENPGVTQLNR
                     LAAHPPFASWRNSEEARTDRPSQQLRSLNGEWRLMRYFLLTHLCGISHRIWCRSLAMSS
                     TSL"
     primer_bind     939..961
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     953..970
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     954..970
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    971..1027
                     /label=MCS
                     /note="pUC18/19 multiple cloning site"
     primer_bind     complement(1040..1056)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1040..1056)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1053..1075)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1064..1080
                     /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(1088..1094,1095..1112,1113..1118))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1133..1154
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     CDS             1465..1656
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     complement(1650..1672)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     misc_feature    1758..1897
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     1813..1832
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1912..1929)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2083..2671)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2163..2182)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(2702..2741)
                     /label=fd terminator
                     /note="central terminator from bacteriophage fd (Otsuka and
                     Kunisawa, 1982)"
     CDS             complement(join(2747..3130,1..432))
                     /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(2747..3130,1..432))
                     /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 atcgctgtta aaaggacaat tacaaacagg aatcgaatgc aaccggcgca ggaacactgc
       61 cagcgcatca acaatatttt cacctgaatc aggatattct tctaatacct ggaatgctgt
      121 tttcccgggg atcgcagtgg tgagtaacca tgcatcatca ggagtacgga taaaatgctt
      181 gatggtcgga agaggcataa attccgtcag ccagtttagt ctgaccatct catctgtaac
      241 atcattggca acgctacctt tgccatgttt cagaaacaac tctggcgcat cgggcttccc
      301 atacaatcga tagattgtcg cacctgattg cccgacatta tcgcgagccc atttataccc
      361 atataaatca gcatccatgt tggaatttaa tcgcggcctg gagcaagacg tttcccgttg
      421 aatatggctc ataacacccc ttgtattact gtttatgtaa gcagacagtt ttattgttca
      481 tgatgatata tttttatctt gtgcaatgta acatcagaga ttttgagaca caacgtggct
      541 ttgttgaata aatcgaactt ttgctgagtt gaaggatcag ctcgagtgcc acctgacgtc
      601 taagaaacca ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcagaat
      661 ttcagataaa aaaaatcctt agctttcgct aaggatgatt tctggaattc gcggccgctt
      721 ctagagacta gtggaagaca tcgctagaga cctgcaccat atgcggtgtg aaataccgca
      781 cagatgcgta aggagaaaat accgcatcag gcgccattcg ccattcaggc tgcgcaactg
      841 ttgggaaggg cgatcggtgc gggcctcttc gctattacgc cagctggcga aagggggatg
      901 tgctgcaagg cgattaagtt gggtaacgcc agggttttcc cagtcacgac gttgtaaaac
      961 gacggccagt gaattcgagc tcggtacccg gggatcctct agagtcgacc tgcaggcatg
     1021 caagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa
     1081 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga
     1141 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt
     1201 gccagctgca ttaatgaatc ggccaacgcg cgggggtctc ttgccatgtc ttctaacacc
     1261 tacatctgta ttaacgaagc gctggcattg accctgagtg atttttctct ggtcccgccg
     1321 catccatacc gccagttgtt taccctcaca acgttccagt aaccgggcat gttcatcatc
     1381 agtaacccgt atcgtgagca tcctctctcg tttcatcggt atcattaccc catgaacaga
     1441 aatccccctt acacggaggc atcagtgacc aaacaggaaa aaaccgccct taacatggcc
     1501 cgctttatca gaagccagac attaacgctt ctggagaaac tcaacgagct ggacgcggat
     1561 gaacaggcag acatctgtga atcgcttcac gaccacgctg atgagcttta ccgcagctgc
     1621 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggcgacggtc
     1681 acagcttgtc tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt
     1741 gttggcgggt gtcggggcgc agccatgacc cagtcacgta gcgatagcgg agtgtatact
     1801 ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccattgc ggtgtgaaat
     1861 accgcacaga tgcgtaagga gaaaataccg catcaggcgc tcttccgctt cctcgctcac
     1921 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     1981 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     2041 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     2101 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     2161 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     2221 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     2281 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     2341 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     2401 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     2461 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     2521 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     2581 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     2641 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctaccggtcc
     2701 gtccaaaaaa aaaggctcca aaaggagcct ttaattgtat cggtgattag aaaaactcat
     2761 cgagcatcaa atgaaactgc aatttattca tatcaggatt atcaatacca tatttttgaa
     2821 aaagccgttt ctgtaatgaa ggagaaaact caccgaggca gttccatagg atggcaagat
     2881 cctggtatcg gtctgcgatt ccgactcgtc caacatcaat acaacctatt aatttcccct
     2941 cgtcaaaaat aaggttatca agtgagaaat caccatgagt gacgactgaa tccggtgaga
     3001 atggcaaaag cttatgcatt tctttccaga cttgttcaac aggccagcca ttacgctcgt
     3061 catcaaaatc actcgcatca accaaaccgt tattcattcg tgattgcgcc tgagcaagac
     3121 gaaatacgcg
//
