LOCUS       sequence-12471-e        8746 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8746)
  TITLE     New vectors Aug. 2010
REFERENCE   2  (bases 1 to 8746)
  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"
COMMENT     Sequence Label: MDC1 in pENTR4 (330-5)
FEATURES             Location/Qualifiers
     source          1..8746
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      106..149
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      281..308
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     primer_bind     316..339
                     /label=pENTR-F
                     /note="pENTR vectors, forward primer"
     protein_bind    358..457
                     /gene="mutant version of attL"
                     /label=attL1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    complement(6975..7074)
                     /gene="mutant version of attL"
                     /label=attL2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(7186..7205)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             7197..8006
                     /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="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPGDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(7264..7283)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      8096..8684
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     8585..8604
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgctagca tggatctcgg
       61 ggacgtctaa ctactaagcg agagtaggga actgccaggc atcaaataaa acgaaaggct
      121 cagtcttccg actgagcctt tcgttttatc tgttgtttgt cggtgaacgc tctcctgagt
      181 aggacaaatc cgccgggagc ggatttgaac gttgtgaagc aacggcccgg agggtggcgg
      241 gcaggacgcc cgccataaac tgccaggcat caaactaagc agaaggccat cctgacggat
      301 ggcctttttg cgtttctaca aactcttcct gttagttagt tacttaagct cgggccccaa
      361 ataatgattt tattttgact gatagtgacc tgttcgttgc aacaaattga taagcaatgc
      421 ttttttataa tgccaacttt gtacaaaaaa gcaggctcca ccatgggaac caattcagtc
      481 gactagtgat atcatggagg acacccaggc tattgactgg gatgttgaag aagaggagga
      541 gacagagcaa tccagtgaat ccttgaggtg taacgtggag ccagtagggc ggctacatat
      601 ctttagtggt gcccatggac cagaaaaaga tttcccacta cacctcggga agaatgtggt
      661 aggccgaatg cctgactgct ctgtggccct gccctttcca tctatctcca aacaacatgc
      721 agagattgaa atcttagcct gggacaaggc acctatcctc cgagactgtg ggagccttaa
      781 tggtactcaa atcctgagac ctcctaaggt tttgagccct ggggtgagtc accgtctgag
      841 ggaccaggaa ttgattctct ttgctgactt gctctgccag taccatcgcc tggatgtctc
      901 tctgcccttt gtctcccggg gccctctgac agtagaagag acacccagag tacagggaga
      961 aactcaaccc cagaggcttc tgttggctga ggactcggag gaggaagtag attttctttc
     1021 tgaaaggcgt atggtaaaaa aatcaaggac cacatcttcc tctgtgatag ttccagagag
     1081 tgatgaagag gggcattccc cggtcctggg cggccttggg ccgccttttg ccttcaattt
     1141 gaacagtgac acagatgtgg aagaaggtca gcaaccagcc acagaggagg cctcctcagc
     1201 tgccagaaga ggtgccactg tagaggcaaa gcagtctgaa gctgaagttg taactgaaat
     1261 ccagcttgaa aaggatcagc ctttagtgaa ggagagggac aatgatacaa aagtcaagag
     1321 gggtgcaggg aatggggtgg ttccagctgg ggtgattctg gagaggagcc aacctcctgg
     1381 agaggacagt gacacagatg tggatgatga cagcaggcct cctggaaggc cagctgaggt
     1441 ccatttggaa agggctcagc cttttggctt catcgacagc gacactgatg cggaagaaga
     1501 gaggatccca gcaaccccag ttgtcattcc tatgaagaag aggaagatct tccatggagt
     1561 aggtacaagg ggtcctggag caccaggcct ggcccatctg caggagagcc aggctggtag
     1621 tgatacagat gtggaagaag gcaaggcccc acaggctgtc cctctggaga aaagccaagc
     1681 ttccatggtt atcaacagcg atacagatga cgaggaagaa gtctcagcag cgctgacttt
     1741 ggcacatctg aaagagagcc agcctgctat atggaacaga gatgcagaag aggacatgcc
     1801 ccaacgtgtg gtccttctgc agcgaagcca aaccaccact gagagagaca gtgacacaga
     1861 cgtggaggag gaagagctcc cagtggaaaa tagagaagct gtcctcaagg atcacacaaa
     1921 gattagagcc cttgttagag cacattcaga aaaggaccaa cctccttttg gggacagtga
     1981 tgacagtgtg gaagcagata agagctcacc tgggatccac ctggagagaa gccaagcctc
     2041 caccacagtg gacatcaaca cacaagtgga gaaggaagtc ccgccagggt cagccattat
     2101 gcatataaag aagcatcagg tgtctgtgga ggggacaaat caaacagatg tgaaagcagt
     2161 tgggggacca gcaaagctgc ttgtggtatc tctagaggaa gcctggcctc tgcatgggga
     2221 ctgtgaaaca gatgcagagg agggcacctc cctaacagcc tcagtagttg cagatgtaag
     2281 aaagagccag cttccagcag aaggggatgc tggggcagag tgggctgcag ctgttcttaa
     2341 gcaggagaga gctcatgagg tgggggccca gggtgggcca cctgtggcac aagtggagca
     2401 ggacctccct atctcaagag agaacctcac agatctggtg gtggacacag acactctagg
     2461 ggaatccacc cagccacaga gagagggagc ccaggtcccc acaggaaggg agagagaaca
     2521 acatgtgggt gggaccaagg actctgaaga caactatggt gattctgaag atctggacct
     2581 acaagctacc cagtgctttc tggagaatca gggcctggaa gcagtccaga gcatggagga
     2641 tgaacctacc caggccttca tgttgactcc accccaagag cttggccctt cccattgcag
     2701 cttccagaca acaggtaccc tagatgaacc atgggaggtc ctggctacac agccattctg
     2761 tctgagagag tctgaggact ctgagaccca gccttttgac acgcaccttg aggcctatgg
     2821 accttgcctg tctccaccta gggcaatacc aggagaccaa catccagaga gcccagttca
     2881 cacagagcca atggggattc aaggcagagg gaggcagact gtggataaag tcatgggtat
     2941 accaaaagaa acagcagaga gggtgggccc tgagagaggg ccattggaga gagaaactga
     3001 gaaactgcta ccagaaagac agacagatgt gacaggagag gaagaattaa ccaaggggaa
     3061 acaggacaga gaacaaaaac agttgttagc tagagacacc cagagacaag aatctgacaa
     3121 aaatggggaa agtgcaagtc ctgaaagaga tagggagagt ttgaaggtag aaattgagac
     3181 atctgaggaa atacaagaga aacaagtaca gaagcagacc cttccaagca aagcatttga
     3241 gagagaagta gagagaccag tagcaaacag agagtgcgat ccagccgagt tagaagagaa
     3301 ggtgcccaaa gtgatcctgg agagagatac acagagaggg gagccagagg gagggagcca
     3361 ggaccagaaa gggcaggcct ccagcccaac accagagcct ggggtggggg cgggggacct
     3421 tccgggacct acctcagccc ccgtaccttc tgggagccag tcaggtggaa ggggatcccc
     3481 agtgagcccc aggaggcatc agaaaggcct cctgaattgc aagatgccac ctgctgagaa
     3541 ggcttccagg atcagagctg ctgagaaggt ttccaggggc gatcaggaat ctccagatgc
     3601 ttgtctgcct cctgcagtac ctgaagcccc agccccaccc caaaagcccc ttaactctca
     3661 gagccagaaa catcttgcac ctccgcccct tctttctccc cttttacctt ctatcaagcc
     3721 aaccgttcgt aagaccaggc aagatgggag tcaggaagct ccagaggctc ccttgtcctc
     3781 agagctggag cctttccacc caaagcctaa aattagaact cggaagtcct ccagaatgac
     3841 accctttcca gctacctctg ctgcccctga gccccaccct tccacctcca cagcccagcc
     3901 agtcactccc aagcccacat ctcaggccac taggagcagg acaaataggt cctctgtcaa
     3961 gacccctgaa ccagttgtcc ccacagcccc tgagctccag ccttccacct ccacagacca
     4021 gcctgtcacc tctgagccca catctcaggt tactagggga agaaaaagta gatcctctgt
     4081 caagacccct gaaacagttg tgcccacagc ccttgagctc cagccttcca cctccaccga
     4141 ccgacctgtc acctctgaac ccacctctca ggctactagg ggaagaaaaa atagatcctc
     4201 tgtcaagacc cctgaaccag ttgtccccac agcccctgag ctccagcctt ccacctccac
     4261 agaccagcct gtcacttctg agcccacata tcaggctact aggggaagaa aaaatagatc
     4321 ctctgtcaag acccctgaac cagttgtgcc cacagcccct gagctccggc cttccacctc
     4381 cacagaccga cctgtcaccc ccaagcccac atctcggacc actaggagca ggacaaatat
     4441 gtcctctgtc aagacccctg aaacagttgt ccccacagcc cctgagctcc agatttccac
     4501 ctccacagac caacctgtca cccctaagcc cacatctcgg accactagga gcaggacaaa
     4561 tatgtcctct gtgaagaacc ctgaatcaac tgtccctata gcccctgagc tcccaccttc
     4621 cacctccaca gagcagcctg tcacccctga gcccacatct cgggctacta ggggaagaaa
     4681 aaatagatcc tctggcaaga cccctgaaac acttgtcccc acagccccta agctcgagcc
     4741 ttccacttcc acagaccaac ctgtcactcc tgagcccaca tctcaggcca ccaggggcag
     4801 gacaaatagg tcctctgtga agacccctga aacagttgtc cccacagccc ctgagctcca
     4861 gccttccacc tccacagacc agcctgttac ccctgagcct acgtctcagg ctactagggg
     4921 aagaacagat agatcctctg tcaagactcc tgaaacagtt gtccccacag cccctgagct
     4981 acaggcttcc gcctccacag accagcctgt cacctctgag cccacatctc ggaccactag
     5041 gggaagaaaa aatcggtcct ctgtcaagac ccctgaaaca gttgtgcccg cagcccctga
     5101 gctccagcct cccacctcca cagaccgacc tgtcacccct gagcccacat ctcgggccac
     5161 taggggcagg acaaataggt cctctgtcaa gacccctgaa tcaattgtcc ctatagcccc
     5221 tgagcttcag ccttccacct ccagaaacca gcttgtcacc cctgagccca catctcgggc
     5281 cactaggtgc aggacaaata ggtcctctgt caagacccct gagccagttg tccccacagc
     5341 ccctgagccc catcctacca cctccacaga ccagcctgtc acccccaagc tcacatctag
     5401 ggccactagg agaaagacaa ataggtcctc tgtcaagact cccaaaccag ttgaaccagc
     5461 agcctctgat cttgagcctt ttacccccac agaccagtcc gtcacccctg aggccatagc
     5521 tcagggtggt cagagcaaaa cactgaggtc ttccacagta agagctatgc cggttcctac
     5581 cacccctgaa ttccaatctc ctgtcaccac agaccagcct atttcccctg agcctattac
     5641 tcaacccagt tgcatcaaga ggcagagagc cgctgggaac cctggctccc tcgcagctcc
     5701 cattgaccat aagccttgct ctgcaccctt ggaacctaaa tcccaggcct caaggaacca
     5761 aagatgggga gcagtgagag cagctgaatc ccttacagcc attcctgagc ctgcctctcc
     5821 ccagcttctt gagacaccaa ttcatgcctc ccagatccaa aaggtggaac cagcaggtag
     5881 atctaggttc accccggagc tccagcctaa ggcctctcaa agccgcaaga ggtctttagc
     5941 taccatggat tcaccaccac atcaaaaaca gccccaaaga ggggaagtct cccagaagac
     6001 agtgattatc aaggaagagg aagaagatac tgcagagaag ccagggaagg aagaggatgt
     6061 cgtgactcca aaaccaggca agagaaagag agaccaggca gaggaggagc ccaacagaat
     6121 accaagccgc agcctccgac ggaccaaact taaccaagaa tcaacagccc ccaaagtgct
     6181 cttcacagga gtggtggatg ctcggggaga gcgggctgtg ctggcactgg ggggaagtct
     6241 ggctggttca gcggcagagg cttcccacct ggtcactgat cgcatccgcc ggacagtcaa
     6301 gttcctgtgt gccctggggc ggggaatccc cattctgtcc ctggactggc tgcatcagtc
     6361 ccgcaaggct ggtttcttct tacccccgga tgaatatgtg gtgaccgacc ctgagcaaga
     6421 gaagaacttt ggctttagcc ttcaagacgc actgagcagg gctcgggagc gaaggctgct
     6481 agagggctat gagatctatg tgacccctgg agtccagcca ccaccacctc agatgggaga
     6541 gattattagc tgctgtggag gcacatacct acccagcatg cctcggtcct ataagcctca
     6601 gagagttgtg atcacatgcc ctcaggactt ccctcattgc tccattccac tacgggttgg
     6661 gctgcccctc ctctcgcctg agttcctgct gactggagtg ctgaagcagg aagccaagcc
     6721 agaggccttt gtcctctccc ctttggagat gtcatccacc tgagaactcc actacccttt
     6781 tccctcccag accacgaatt agaagatatg tggaagaaag aactcagggc gttagaaagg
     6841 attggggtat attgatacaa cttgtcctgg aacatgggtg ggaccagaaa tctttatgaa
     6901 taaatgaaaa gataagggat ttggaagcca caggttgttt tttgtttgtt tgtttgtttt
     6961 tttaatggat ctagacccag ctttcttgta caaagttggc attataagaa agcattgctt
     7021 atcaatttgt tgcaacgaac aggtcactat cagtcaaaat aaaatcatta tttgccatcc
     7081 agctgcagct ctggcccgtg tctcaaaatc tctgatgtta cattgcacaa gataaaaata
     7141 tatcatcatg aacaataaaa ctgtctgctt acataaacag taatacaagg ggtgttatga
     7201 gccatattca acgggaaacg tcgaggccgc gattaaattc caacatggat gctgatttat
     7261 atgggtataa atgggctcgc gataatgtcg ggcaatcagg tgcgacaatc tatcgcttgt
     7321 atgggaagcc cgatgcgcca gagttgtttc tgaaacatgg caaaggtagc gttgccaatg
     7381 atgttacaga tgagatggtc agactaaact ggctgacgga atttatgcct cttccgacca
     7441 tcaagcattt tatccgtact cctggtgatg catggttact caccactgcg atccccggaa
     7501 aaacagcatt ccaggtatta gaagaatatc ctgattcagg tgaaaatatt gttgatgcgc
     7561 tggcagtgtt cctgcgccgg ttgcattcga ttcctgtttg taattgtcct tttaacagcg
     7621 atcgcgtatt tcgtctcgct caggcgcaat cacgaatgaa taacggtttg gttgatgcga
     7681 gtgattttga tgacgagcgt aatggctggc ctgttgaaca agtctggaaa gaaatgcata
     7741 aacttttgcc attctcaccg gattcagtcg tcactcatgg tgatttctca cttgataacc
     7801 ttatttttga cgaggggaaa ttaataggtt gtattgatgt tggacgagtc ggaatcgcag
     7861 accgatacca ggatcttgcc atcctatgga actgcctcgg tgagttttct ccttcattac
     7921 agaaacggct ttttcaaaaa tatggtattg ataatcctga tatgaataaa ttgcagtttc
     7981 atttgatgct cgatgagttt ttctaatcag aattggttaa ttggttgtaa cattattcag
     8041 attgggcccc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     8101 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     8161 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     8221 gagcgcagat accaaatact gttcttctag tgtagccgta gttaggccac cacttcaaga
     8281 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     8341 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     8401 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     8461 ccgaactgag atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa
     8521 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     8581 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     8641 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     8701 cctttttacg gttcctggcc ttttgctggc cttttgctca catgtt
//
