LOCUS       sequence-267199-        3634 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 3634)
  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 3634)
  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..3634
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     92..109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     terminator      complement(387..473)
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     primer_bind     522..544
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     536..553
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    593..688
                     /gene="mutant version of attL"
                     /label=attL4
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     protein_bind    1680..1803
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1875..1894)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1876..1894)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1899..1915)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1899..1915)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2017..2036)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2028..2837
                     /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(2095..2114)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      2984..3572
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3473..3492
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccgc gttaacgcta ccatggagct ccaaataatg
      601 attttatttt gactgatagt gacctgttcg ttgcaacaaa ttgataagca atgctttttt
      661 ataatgccaa ctttgtatag aaaagttggg ctgctgattc atatcccagt gaggtcatct
      721 gaattttggg ggacctcaaa tagaaatcat atgggggaaa gaatgagcct agctcaaacc
      781 agacaggctg agagcttatc ttgccaggac ataggctccc cggtgggtgg gggagatggc
      841 cattgctgcc tttctttcct tccccactga atttaagcag atgcagttag tcactgtgca
      901 aaggagtatg tgcaaatcta agtattcatg tttatgtgtg gttgtcaggt ctaaatcagc
      961 caggcagtaa tttctctggg agcccaatgt aggcactaaa ttttgcactc aaccaccgaa
     1021 tggtccacat gccccttaat ctgtcttcct gcaggagggc tcccttggca ccatcctgcc
     1081 tgaacttggg gccagccttt cactggagcc ccttcccgtg aattatggcc tacagcatgg
     1141 gggctgaata gaaaaaaaaa aaccctccag aaattcaact ggaagggctg accggaattc
     1201 aacctaatgt tgagaatagc aggaaagctt ctattattct acttcattct atctgctatg
     1261 aggcagtgcg gtcgcctgga gtcgactgag cttaactcat tacgctggat tacggagtca
     1321 cagccatggt gcctgctaaa gccaaatatc tcactcctga gactcactac acttcctgat
     1381 aggatgcact tgcgtgtctc ctagtgacct taaacactaa taaaggagtc ataatatgac
     1441 ctagacaaga cctaggagcc aatcagaatg caaatgattg atgggagtgc ttttggttat
     1501 ctactgtttg tcctgacgct tggcgtggtt ctttgtatta tgattgtgtg ggcagtaggc
     1561 ttaaaaatac atacaggcat tgctttccac attgttgaag gttaatccta aggtgtggca
     1621 aagaaactat attttataga ctggaacctc atgggcacca gctagttttt gctcttcctc
     1681 aagtttgtac aaaaaagttg aacgagaaac gtaaaatgat ataaatatca atatattaaa
     1741 ttagattttg cataaaaaac agactacata atactgtaaa acacaacata tgcagtcact
     1801 atgaatcaac tacttagatg gtattagtga cctgtagaat tcgagctcta gagctgcagg
     1861 gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt tcctggcagc
     1921 tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat atatcatcat
     1981 gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg agccatattc
     2041 aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta tatgggtata
     2101 aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg tatgggaagc
     2161 ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat gatgttacag
     2221 atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc atcaagcatt
     2281 ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga aaaacagcat
     2341 tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg ctggcagtgt
     2401 tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc gatcgcgtat
     2461 ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg agtgattttg
     2521 atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat aaacttttgc
     2581 cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac cttatttttg
     2641 acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca gaccgatacc
     2701 aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta cagaaacggc
     2761 tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt catttgatgc
     2821 tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag agcattacgc
     2881 tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt tacgcgtcgt
     2941 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3001 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3061 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3121 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3181 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3241 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3301 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3361 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3421 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3481 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     3541 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     3601 tcctggcctt ttgctggcct tttgctcaca tgtt
//
