LOCUS       sequence-272006-        3635 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone contains human modified 161k 
            enhancer sequence. with NR2F2 binding sites mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3635)
  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 3635)
  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: p5E-161K enhancer-NR2F2-MUT
FEATURES             Location/Qualifiers
     source          1..3635
                     /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    597..692
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1685..1808
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1880..1899)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1881..1899)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1904..1920)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1904..1920)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2022..2041)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2033..2842
                     /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(2100..2119)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2989..3577
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3478..3497
                     /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 ccgagttaac gctaccatgg agctccaaat
      601 aatgatttta ttttgactga tagtgacctg ttcgttgcaa caaattgata agcaatgctt
      661 ttttataatg ccaactttgt atagaaaagt tgggctgctg attcatatcc cagtgaggtc
      721 atctgaattt tgggggacct caaatagaaa tcatatgggg gaaagaatga gcctagctca
      781 aaccagacag gctgagagct tatcttgcca ggacataggc tccccggtgg gtgggggaga
      841 tggccattgc tgcctttctt tccttcccca ctgaatttaa gcagatgcag ttagtcactg
      901 tgcaaaggag tatgtgcaaa tctaagtatt catgtttatg tgtggttgtc aggtctaaat
      961 cagccaggca gtaatttctc tgggagccca atgtaggcac taaattttgc actcaaccac
     1021 cgaatggtcc acatgcccct taatctgtct tcctgcagga gggctccctt ggcaccatcc
     1081 tgcctgaact tggggccagc ctttcactgg agccccttcc cgtgaattat ggcctacagc
     1141 atgggggctg aatagaaaaa aaaaaaaccc tccagaaatt caactggaag ggctgaccgg
     1201 aattcaacct aatgttgaga atagcaggaa agcttctatt attctacttc attctatctg
     1261 ctatgaggca gtgcggtcgc ctggagtcga ctgagcttaa ctcatttccc tggatttccg
     1321 agtcacagcc atggtgcctg ctatagttaa cgatcgaact cctgagactc actacacttc
     1381 ctgataggac gtactcacgt accgcctagt aacatcatac gttcctatag gagtcataat
     1441 atgacctaga caagacctag gagccaatca gaatgcaaat gattgatggg agtgcttttg
     1501 gttatctact gtttgtcctg acgcttggcg tggttctttg tattatgatt gtgtgggcag
     1561 taggcttaaa aatacataca ggcattgctt tccacattgt tgaaggttaa tcctaaggtg
     1621 tggcaaagaa actatatttt atagactgga acctcatggg caccagctag tttttgctct
     1681 tcctcaagtt tgtacaaaaa agttgaacga gaaacgtaaa atgatataaa tatcaatata
     1741 ttaaattaga ttttgcataa aaaacagact acataatact gtaaaacaca acatatgcag
     1801 tcactatgaa ccaactactt agatggtatt agtgacctgt agaattcgag ctctagagct
     1861 gcagggcggc cgcgatatcc cctatagtga gtcgtattac atggtcatag ctgtttcctg
     1921 gcagctctgg cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc
     1981 atcatgaaca ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca
     2041 tattcaacgg gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg
     2101 gtataaatgg gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg
     2161 gaagcccgat gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt
     2221 tacagatgag atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa
     2281 gcattttatc cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac
     2341 agcattccag gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc
     2401 agtgttcctg cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg
     2461 cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga
     2521 ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact
     2581 tttgccattc tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat
     2641 ttttgacgag gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg
     2701 ataccaggat cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa
     2761 acggcttttt caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt
     2821 gatgctcgat gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat
     2881 tacgctgact tgacgggacg gcgcaagctc atgaccaaaa tcccttaacg tgagttacgc
     2941 gtcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     3001 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     3061 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     3121 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     3181 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     3241 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     3301 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     3361 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     3421 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     3481 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     3541 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     3601 acggttcctg gccttttgct ggccttttgc tcaca
//
