LOCUS       sequence-336489-        4015 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  PCR template for reporter gene with mouse Rhodopsin promoter with 
            DsRed.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4015)
  AUTHORS   White MA, Myers CA, Corbo JC, Cohen BA
  TITLE     Massively parallel in vivo enhancer assay reveals that highly local 
            features determine the cis-regulatory function of ChIP-seq peaks.
  JOURNAL   Proc Natl Acad Sci U S A. 2013 Jul 16;110(29):11952-7. doi: 
            10.1073/pnas.1307449110. Epub 2013 Jul 1.
  PUBMED    23818646
REFERENCE   2  (bases 1 to 4015)
  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"; doi: 
            "10.1073/pnas.1307449110"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2013-07-16- 16"; volume: "110"; issue: "29"; pages: "11952-7"
COMMENT     Sequence Label: pJK01_Rho_minprox_DsREd
FEATURES             Location/Qualifiers
     source          1..4015
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      251..260
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             257..934
                     /codon_start=1
                     /product="improved tetrameric variant of DsRed fluorescent 
                     protein"
                     /label=DsRed2
                     /note="mammalian codon-optimized"
                     /translation="MASSENVITEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEGHNTVK
                     LKVTKGGPLPFAWDILSPQFQYGSKVYVKHPADIPDYKKLSFPEGFKWERVMNFEDGGV
                     ATVTQDSSLQDGCFIYKVKFIGVNFPSDGPVMQKKTMGWEASTERLYPRDGVLKGETHK
                     ALKLKDGGHYLVEFKSIYMAKKPVQLPGYYYVDAKLDITSHNEDYTIVEQYERTEGRHH
                     LFL"
     primer_bind     complement(437..457)
                     /label=DsRed1-N
                     /note="DsRed1, reverse primer"
     primer_bind     849..872
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(1039..1058)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1104..1159
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1158..1177)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1518..1534)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1518..1534)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1531..1553)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1542..1558
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(1566..1572,1573..1590,1591..1596))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1611..1632
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1691..1886
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1737..1872
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1892..2026
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1929..1948)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1983..2002
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2094..2111)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2265..2853)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2345..2364)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3024..3815,3816..3884))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     3647..3666
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3885..3989)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacatat ctctagaatg tcaccttggc ccctctgcaa gccaattagg ccccggtggc
       61 agcagtggga ttagcgttag tatgatatct cgcggatgct gaatcagcct ctggcttagg
      121 gagagaaggt cactttataa gggtctgggg ggggtcagtg cctggagttg cgctgtggga
      181 gccgtcagag gctgagctcg ccaagcagcc ttggtctctg tcggtaccgc gggcccggga
      241 tccaccggtc gccaccatgg cctcctccga gaacgtcatc accgagttca tgcgcttcaa
      301 ggtgcgcatg gagggcaccg tgaacggcca cgagttcgag atcgagggcg agggcgaggg
      361 ccgcccctac gagggccaca acaccgtgaa gctgaaggtg accaagggcg gccccctgcc
      421 cttcgcctgg gacatcctgt ccccccagtt ccagtacggc tccaaggtgt acgtgaagca
      481 ccccgccgac atccccgact acaagaagct gtccttcccc gagggcttca agtgggagcg
      541 cgtgatgaac ttcgaggacg gcggcgtggc gaccgtgacc caggactcct ccctgcagga
      601 cggctgcttc atctacaagg tgaagttcat cggcgtgaac ttcccctccg acggccccgt
      661 gatgcagaag aagaccatgg gctgggaggc ctccaccgag cgcctgtacc cccgcgacgg
      721 cgtgctgaag ggcgagaccc acaaggccct gaagctgaag gacggcggcc actacctggt
      781 ggagttcaag tccatctaca tggccaagaa gcccgtgcag ctgcccggct actactacgt
      841 ggacgccaag ctggacatca cctcccacaa cgaggactac accatcgtgg agcagtacga
      901 gcgcaccgag ggccgccacc acctgttcct gtagcggccg ttactcgaga tcgatggccg
      961 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1021 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1081 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1141 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     1201 gaatgagtat ttggtttaga gtttggcaac atatgccata tgctggctgc catgaacaaa
     1261 ggtggctata aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc
     1321 atagaaaagc cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt
     1381 ctttaacatc cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct
     1441 gactactccc agtcatagct gtccctcttc tcttatgaag atccctcgac ctgcagccca
     1501 agcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     1561 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     1621 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     1681 cagcggatcc gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     1741 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     1801 tttatgcaga ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct
     1861 tttttggagg cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta
     1921 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     1981 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga
     2041 atcggccaac gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc
     2101 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     2161 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     2221 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     2281 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     2341 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     2401 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcaa
     2461 tgctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     2521 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     2581 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     2641 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact
     2701 agaaggacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     2761 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     2821 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     2881 tctgacgctc agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa
     2941 aggatcttca cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata
     3001 tatgagtaaa cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg
     3061 atctgtctat ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata
     3121 cgggagggct taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg
     3181 gctccagatt tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct
     3241 gcaactttat ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt
     3301 tcgccagtta atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc
     3361 tcgtcgtttg gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga
     3421 tcccccatgt tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt
     3481 aagttggccg cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc
     3541 atgccatccg taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa
     3601 tagtgtatgc ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca
     3661 catagcagaa ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca
     3721 aggatcttac cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct
     3781 tcagcatctt ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc
     3841 gcaaaaaagg gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa
     3901 tattattgaa gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt
     3961 tagaaaaata aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
