LOCUS       sequence-272019-        3877 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 3' entry clone contains puromycin driven by 
            simian virus 40 (SV40).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3877)
  AUTHORS   Sissaoui S, Yu J, Yan A, Li R, Zhu LJ, Kucukural A, Yukselen O, 
            Lawson ND
  TITLE     Genomic Characterization of Endothelial Enhancers Reveals a 
            Multifunctional Role for NR2F2 in Regulation of Arteriovenous Gene 
            Expression.
  JOURNAL   Circ Res. 2020 Feb 17. doi: 10.1161/CIRCRESAHA.119.316075.
  PUBMED    32065070
REFERENCE   2  (bases 1 to 3877)
  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: "Circ Res. 
            2020 Feb 17. doi: 10.1161/CIRCRESAHA.119.316075."
COMMENT     Sequence Label: p3E-SV40-puromycin
FEATURES             Location/Qualifiers
     source          1..3877
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(272..299)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(391..477)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     526..548
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     540..557
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     541..557
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(628..751)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        752..1081
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      932..1067
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             1091..1690
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(1091..1110)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     1587..1607
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     protein_bind    complement(1990..2085)
                     /gene="mutant version of attL"
                     /label=attL3
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(2122..2141)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(2123..2141)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2146..2162)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2146..2162)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2264..2283)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2275..3084
                     /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
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(2342..2361)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3231..3819
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3720..3739
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
       61 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      121 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct
      181 ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatacgcgta
      241 ccgctagcca ggaagagttt gtagaaacgc aaaaaggcca tccgtcagga tggccttctg
      301 cttagtttga tgcctggcag tttatggcgg gcgtcctgcc cgccaccctc cgggccgttg
      361 cttcacaacg ttcaaatccg ctcccggcgg atttgtccta ctcaggagag cgttcaccga
      421 caaacaacag ataaaacgaa aggcccagtc ttccgactga gcctttcgtt ttatttgatg
      481 cctggcagtt ccctactctc gcgttaacgc tagcatggat gttttcccag tcacgacgtt
      541 gtaaaacgac ggccagtctt aagctcgggc cctgcagctc tagagctcga attctacagg
      601 tcactaatac catctaagta gttggttcat agtgactgca tatgttgtgt tttacagtat
      661 tatgtagtct gttttttatg caaaatctaa tttaatatat tgatatttat atcattttac
      721 gtttctcgtt caactttctt gtacaaagtg ggtgtgtcag ttagggtgtg gaaagtcccc
      781 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
      841 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
      901 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
      961 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     1021 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     1081 aaagcttacc atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc
     1141 cagggccgta cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt
     1201 cgatccggac cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt
     1261 cgggctcgac atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac
     1321 cacgccggag agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga
     1381 gttgagcggt tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg
     1441 gcccaaggag cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa
     1501 gggtctgggc agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc
     1561 cgccttcctg gagacctccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac
     1621 cgtcaccgcc gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc
     1681 cggtgcctga cgcccgcccc acgacccgca gcgcccgacc gaaaggagcg cacgacccca
     1741 tgcatcgata aaataaaaga ttttatttag tctccagaaa aaggggggaa tgaaagaccc
     1801 cacctgtagg tttggcaagc tagcttaagt aacgccattt tgcaaggcat ggaaaaatac
     1861 ataactgaga atagagaagt tcagatcaag gtcaggaaca gatggaacag ctgaatatgg
     1921 gccaaacagg atatctgtgg taagcagttc ctgccccggc tcagggccaa gaacagatgg
     1981 aacagctgac aactttatta tacaaagttg gcattataaa aaagcattgc ttatcaattt
     2041 gttgcaacga acaggtcact atcagtcaaa ataaaatcat tatttggagc tccatggtag
     2101 cgttaacgcg gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     2161 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     2221 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     2281 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2341 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2401 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2461 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2521 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2581 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2641 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2701 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2761 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2821 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2881 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2941 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     3001 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     3061 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     3121 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     3181 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3241 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3301 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3361 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3421 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3481 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3541 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3601 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3661 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3721 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3781 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3841 ttacggttcc tggccttttg ctggcctttt gctcaca
//
