LOCUS       sequence-272008-        3633 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone contains human modified 161k 
            enhancer sequence. with ETS binding sites mutated.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3633)
  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 3633)
  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-ETS-CORE-MUT
FEATURES             Location/Qualifiers
     source          1..3633
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..117
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(276..303)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(395..481)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     530..552
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     544..561
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     545..561
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    601..696
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1687..1810
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1882..1901)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1883..1901)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1906..1922)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1906..1922)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2024..2043)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2035..2844
                     /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(2102..2121)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2991..3579
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3480..3499
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
       61 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
      121 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
      181 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatacg
      241 cgtaccgcta gccaggaaga gtttgtagaa acgcaaaaag gccatccgtc aggatggcct
      301 tctgcttagt ttgatgcctg gcagtttatg gcgggcgtcc tgcccgccac cctccgggcc
      361 gttgcttcac aacgttcaaa tccgctcccg gcggatttgt cctactcagg agagcgttca
      421 ccgacaaaca acagataaaa cgaaaggccc agtcttccga ctgagccttt cgttttattt
      481 gatgcctggc agttccctac tctcgcgtta acgctagcat ggatgttttc ccagtcacga
      541 cgttgtaaaa cgacggccag tcttaagctc gggcccgagt taacgctacc atggagctcc
      601 aaataatgat tttattttga ctgatagtga cctgttcgtt gcaacaaatt gataagcaat
      661 gcttttttat aatgccaact ttgtatagaa aagttgggct gctgattcat atcccagtga
      721 ggtcatctga attttggggg acctcaaata gaaatcatat gggggaaaga atgagcctag
      781 ctcaaaccag acaggctgag agcttatctt gccaggacat aggctccccg gtgggtgggg
      841 gagatggcca ttgctgcctt tctttccttc cccactgaat ttaagcagat gcagttagtc
      901 actgtgcaaa ggagtatgtg caaatctaag tattcatgtt tatgtgtggt tgtcaggtct
      961 aaatcagcca ggcagtaatt tctctgggag cccaatgtag gcactaaatt ttgcactcaa
     1021 ccaccgaatg gtccacatgc cccttaatct gtcttcctgc aggagggctc ccttggcacc
     1081 atcctgcctg aacttggggc cagcctttca ctggagcccc ttcccgtgaa ttatggccta
     1141 cagcatgggg gctgaataga aaaaaaaaac cctccagaaa ttcaactgga agggctgacc
     1201 ggaattcaac ctaatgttga gaatagcagg aaagcttcta ttattctact tcattctatc
     1261 tgctatgagg cagtgcggtc gcctggagtc gactgagctt aactcattac gctggattac
     1321 ggagtcacag ccatggtgcc tgctaaagcc aaatatctca ctcctgagac tcactacact
     1381 tcctgatagg atgcacttgc gtgtctccta gtgaccttaa acactaataa aggagtcata
     1441 atatgaccta gacaagacct aggagccaat cagaatgcaa atgattgatg ggagtgcttt
     1501 tggttatcta ctgtttgtcc tgacgcttgg cgtggttctt tgtattatga ttgtgtgggc
     1561 agtaggctta aaaatacata caggcattgc tttccacatt gttgaaggtt aatcctaagg
     1621 tgtggcaaag aaactatatt ttatagactg gaacctcatg ggcaccagct agtttttgct
     1681 cttcctcaag tttgtacaaa aaagttgaac gagaaacgta aaatgatata aatatcaata
     1741 tattaaatta gattttgcat aaaaaacaga ctacataata ctgtaaaaca caacatatgc
     1801 agtcactatg aaccaactac ttagatggta ttagtgacct gtagaattcg agctctagag
     1861 ctgcagggcg gccgcgatat cccctatagt gagtcgtatt acatggtcat agctgtttcc
     1921 tggcagctct ggcccgtgtc tcaaaatctc tgatgttaca ttgcacaaga taaaaatata
     1981 tcatcatgaa caataaaact gtctgcttac ataaacagta atacaagggg tgttatgagc
     2041 catattcaac gggaaacgtc gaggccgcga ttaaattcca acatggatgc tgatttatat
     2101 gggtataaat gggctcgcga taatgtcggg caatcaggtg cgacaatcta tcgcttgtat
     2161 gggaagcccg atgcgccaga gttgtttctg aaacatggca aaggtagcgt tgccaatgat
     2221 gttacagatg agatggtcag actaaactgg ctgacggaat ttatgcctct tccgaccatc
     2281 aagcatttta tccgtactcc tgatgatgca tggttactca ccactgcgat ccccggaaaa
     2341 acagcattcc aggtattaga agaatatcct gattcaggtg aaaatattgt tgatgcgctg
     2401 gcagtgttcc tgcgccggtt gcattcgatt cctgtttgta attgtccttt taacagcgat
     2461 cgcgtatttc gtctcgctca ggcgcaatca cgaatgaata acggtttggt tgatgcgagt
     2521 gattttgatg acgagcgtaa tggctggcct gttgaacaag tctggaaaga aatgcataaa
     2581 cttttgccat tctcaccgga ttcagtcgtc actcatggtg atttctcact tgataacctt
     2641 atttttgacg aggggaaatt aataggttgt attgatgttg gacgagtcgg aatcgcagac
     2701 cgataccagg atcttgccat cctatggaac tgcctcggtg agttttctcc ttcattacag
     2761 aaacggcttt ttcaaaaata tggtattgat aatcctgata tgaataaatt gcagtttcat
     2821 ttgatgctcg atgagttttt ctaatcagaa ttggttaatt ggttgtaaca ctggcagagc
     2881 attacgctga cttgacggga cggcgcaagc tcatgaccaa aatcccttaa cgtgagttac
     2941 gcgtcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc ttgagatcct
     3001 ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc agcggtggtt
     3061 tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt cagcagagcg
     3121 cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt caagaactct
     3181 gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc tgccagtggc
     3241 gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa ggcgcagcgg
     3301 tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac ctacaccgaa
     3361 ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg gagaaaggcg
     3421 gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga gcttccaggg
     3481 ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact tgagcgtcga
     3541 tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa cgcggccttt
     3601 ttacggttcc tggccttttg ctggcctttt gct
//
