LOCUS       sequence-267259-        3704 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Gateway compatible 5' entry clone contains Cytomegalovirus and SP6 
            dual promoter(CMVSSP6).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3704)
  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 3704)
  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-CMVSP6
FEATURES             Location/Qualifiers
     source          1..3704
                     /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"
     promoter        712..1696
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1731..1749
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1731..1748
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1749..1873
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1945..1964)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1946..1964)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1969..1985)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1969..1985)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2087..2106)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2098..2907
                     /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(2165..2184)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3054..3642
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3543..3562
                     /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 aaaagttgaa ggcctcttcg ctattacgcc agtcgaccgc
      721 caattcaata tggcgtatat ggactcatgc caattcaata tggtggatct ggacctgtgc
      781 caattcaata tggcgtatat ggactcgtgc caattcaata tggtggatct ggaccccagc
      841 caattcaata tggcggactt ggcaccatgc caattcaata tggcggacct ggcactgtgc
      901 caactgggga ggggtctact tggcacggtg ccaagtttga ggaggggtct tggccctgtg
      961 ccaagtccgc catattgaat tggcatggtg ccaataatgg cggccatatt ggctatatgc
     1021 caggatcaat atataggcaa tatccaatat ggccctatgc caatatggct attggccagg
     1081 ttcaatacta tgtattggcc ctatgccata tagtattcca tatatgggtt ttcctattga
     1141 cgtagatagc ccctcccaat gggcggtccc atataccata tatggggctt cctaataccg
     1201 cccatagcca ctcccccatt gacgtcaatg gtctctatat atggtctttc ctattgacgt
     1261 catatgggcg gtcctattga cgtatatggc gcctccccca ttgacgtcaa ttacggtaaa
     1321 tggcccgcct ggctcaatgc ccattgacgt caataggacc acccaccatt gacgtcaatg
     1381 ggatggctca ttgcccattc atatccgttc tcacgccccc tattgacgtc aatgacggta
     1441 aatggcccac ttggcagtac atcaatatct attaatagta acttggcaag tacattacta
     1501 ttggaagtac gccagggtac attggcagta ctcccattga cgtcaatggc ggtaaatggc
     1561 ccgcgatggc tgccaagtac atccccattg acgtcaatgg ggaggggcaa tgacgcaaat
     1621 gggcgttcca ttgacgtaaa tgggcggtag gcgtgcctaa tgggaggtct atataagcaa
     1681 tgctcgttta gggaaccgcc attctgcctg gggacgtcgg agcaagcttg atttaggtga
     1741 cactatagac aagtttgtac aaaaaagttg aacgagaaac gtaaaatgat ataaatatca
     1801 atatattaaa ttagattttg cataaaaaac agactacata atactgtaaa acacaacata
     1861 tgcagtcact atgaaccaac tacttagatg gtattagtga cctgtagaat tcgagctcta
     1921 gagctgcagg gcggccgcga tatcccctat agtgagtcgt attacatggt catagctgtt
     1981 tcctggcagc tctggcccgt gtctcaaaat ctctgatgtt acattgcaca agataaaaat
     2041 atatcatcat gaacaataaa actgtctgct tacataaaca gtaatacaag gggtgttatg
     2101 agccatattc aacgggaaac gtcgaggccg cgattaaatt ccaacatgga tgctgattta
     2161 tatgggtata aatgggctcg cgataatgtc gggcaatcag gtgcgacaat ctatcgcttg
     2221 tatgggaagc ccgatgcgcc agagttgttt ctgaaacatg gcaaaggtag cgttgccaat
     2281 gatgttacag atgagatggt cagactaaac tggctgacgg aatttatgcc tcttccgacc
     2341 atcaagcatt ttatccgtac tcctgatgat gcatggttac tcaccactgc gatccccgga
     2401 aaaacagcat tccaggtatt agaagaatat cctgattcag gtgaaaatat tgttgatgcg
     2461 ctggcagtgt tcctgcgccg gttgcattcg attcctgttt gtaattgtcc ttttaacagc
     2521 gatcgcgtat ttcgtctcgc tcaggcgcaa tcacgaatga ataacggttt ggttgatgcg
     2581 agtgattttg atgacgagcg taatggctgg cctgttgaac aagtctggaa agaaatgcat
     2641 aaacttttgc cattctcacc ggattcagtc gtcactcatg gtgatttctc acttgataac
     2701 cttatttttg acgaggggaa attaataggt tgtattgatg ttggacgagt cggaatcgca
     2761 gaccgatacc aggatcttgc catcctatgg aactgcctcg gtgagttttc tccttcatta
     2821 cagaaacggc tttttcaaaa atatggtatt gataatcctg atatgaataa attgcagttt
     2881 catttgatgc tcgatgagtt tttctaatca gaattggtta attggttgta acactggcag
     2941 agcattacgc tgacttgacg ggacggcgca agctcatgac caaaatccct taacgtgagt
     3001 tacgcgtcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     3061 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     3121 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     3181 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     3241 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     3301 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     3361 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     3421 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     3481 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     3541 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     3601 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     3661 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgtt
//
