LOCUS       sequence-272013-        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     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"
     promoter        716..1700
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     promoter        1735..1753
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1735..1752
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     protein_bind    1753..1877
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(1949..1968)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1950..1968)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1973..1989)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1973..1989)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2091..2110)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             2102..2911
                     /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(2169..2188)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     rep_origin      3058..3646
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3547..3566
                     /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 tgaaggcctc ttcgctatta cgccagtcga
      721 ccgccaattc aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct
      781 gtgccaattc aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc
      841 cagccaattc aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact
      901 gtgccaactg gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc
      961 tgtgccaagt ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat
     1021 atgccaggat caatatatag gcaatatcca atatggccct atgccaatat ggctattggc
     1081 caggttcaat actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta
     1141 ttgacgtaga tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat
     1201 accgcccata gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg
     1261 acgtcatatg ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg
     1321 taaatggccc gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc
     1381 aatgggatgg ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac
     1441 ggtaaatggc ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt
     1501 actattggaa gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa
     1561 tggcccgcga tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc
     1621 aaatgggcgt tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa
     1681 gcaatgctcg tttagggaac cgccattctg cctggggacg tcggagcaag cttgatttag
     1741 gtgacactat agacaagttt gtacaaaaaa gttgaacgag aaacgtaaaa tgatataaat
     1801 atcaatatat taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa
     1861 catatgcagt cactatgaac caactactta gatggtatta gtgacctgta gaattcgagc
     1921 tctagagctg cagggcggcc gcgatatccc ctatagtgag tcgtattaca tggtcatagc
     1981 tgtttcctgg cagctctggc ccgtgtctca aaatctctga tgttacattg cacaagataa
     2041 aaatatatca tcatgaacaa taaaactgtc tgcttacata aacagtaata caaggggtgt
     2101 tatgagccat attcaacggg aaacgtcgag gccgcgatta aattccaaca tggatgctga
     2161 tttatatggg tataaatggg ctcgcgataa tgtcgggcaa tcaggtgcga caatctatcg
     2221 cttgtatggg aagcccgatg cgccagagtt gtttctgaaa catggcaaag gtagcgttgc
     2281 caatgatgtt acagatgaga tggtcagact aaactggctg acggaattta tgcctcttcc
     2341 gaccatcaag cattttatcc gtactcctga tgatgcatgg ttactcacca ctgcgatccc
     2401 cggaaaaaca gcattccagg tattagaaga atatcctgat tcaggtgaaa atattgttga
     2461 tgcgctggca gtgttcctgc gccggttgca ttcgattcct gtttgtaatt gtccttttaa
     2521 cagcgatcgc gtatttcgtc tcgctcaggc gcaatcacga atgaataacg gtttggttga
     2581 tgcgagtgat tttgatgacg agcgtaatgg ctggcctgtt gaacaagtct ggaaagaaat
     2641 gcataaactt ttgccattct caccggattc agtcgtcact catggtgatt tctcacttga
     2701 taaccttatt tttgacgagg ggaaattaat aggttgtatt gatgttggac gagtcggaat
     2761 cgcagaccga taccaggatc ttgccatcct atggaactgc ctcggtgagt tttctccttc
     2821 attacagaaa cggctttttc aaaaatatgg tattgataat cctgatatga ataaattgca
     2881 gtttcatttg atgctcgatg agtttttcta atcagaattg gttaattggt tgtaacactg
     2941 gcagagcatt acgctgactt gacgggacgg cgcaagctca tgaccaaaat cccttaacgt
     3001 gagttacgcg tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     3061 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     3121 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     3181 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     3241 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     3301 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     3361 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     3421 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     3481 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     3541 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     3601 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     3661 ggccttttta cggttcctgg ccttttgctg gccttttgct caca
//
