LOCUS       sequence-491431-        6204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes TdTomato driven by the CAG promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6204)
  AUTHORS   Schick E, McCaffery SD, Keblish EE, Thakurdin C, Emerson MM
  TITLE     Lineage tracing analysis of cone photoreceptor associated 
            cis-regulatory elements in the developing chicken retina.
  JOURNAL   Sci Rep. 2019 Jun 27;9(1):9358. doi: 10.1038/s41598-019-45750-7.
  PUBMED    31249345
REFERENCE   2  (bases 1 to 6204)
  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: "Sci Rep."; 
            date: "2019-06-27"; volume: "9(1):9358. doi"; pages: " 
            10.1038/s41598-019-45750-7"
COMMENT     Sequence Label: CAG::TdTomato
FEATURES             Location/Qualifiers
     source          1..6204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      167..755
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     656..675
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     909..926
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    994..1128
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1018..1037)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1072..1091
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(1134..1329)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1148..1283
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    1388..1409
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(1424..1429,1430..1447,1448..1454)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1462..1478
                     /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)."
     primer_bind     1467..1489
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     1486..1502
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1486..1502
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1845..1864
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1863..1918
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1964..1983
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     primer_bind     2208..2225
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(2422..3123)
                     /codon_start=1
                     /product="dimeric variant of DsRed fluorescent protein 
                     (Shaner et al., 2004)"
                     /label=dTomato
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEVIKEFMRFKVRMEGSMNGHEFEIEGEGEGRPYEGTQTA
                     KLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYKKLSFPEGFKWERVMNFEDGG
                     LVTVTQDSSLQDGTLIYKVKMRGTNFPPDGPVMQKKTMGWEASTERLYPRDGVLKGEIH
                     QALKLKDGGHYLVEFKTIYMAKKPVQLPGYYYVDTKLDITSHNEDYTIVEQYERSEGRH
                     HLFLYGMDELYK"
     primer_bind     complement(2505..2528)
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     primer_bind     complement(2703..2722)
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     primer_bind     2972..2990
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     regulatory      3120..3129
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(3261..3280)
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     intron          3288..4304
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     complement(3342..3364)
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     enhancer        4619..4998
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     misc_feature    5009..5100
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(5041..5060)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    5114..5162
                     /note="synthetic polyadenylation signal"
     promoter        5235..5339
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5340..5408,5409..6200)
                     /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     complement(5558..5577)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
ORIGIN
        1 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
       61 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
      121 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
      181 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
      241 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
      301 taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
      361 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
      421 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
      481 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
      541 gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
      601 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
      661 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
      721 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
      781 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
      841 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
      901 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
      961 tccccgcgcg ttggccgatt cattaatgca gcggatccag acatgataag atacattgat
     1021 gagtttggac aaaccacaac tagaatgcag tgaaaaaaat gctttatttg tgaaatttgt
     1081 gatgctattg ctttatttgt aaccattata agctgcaata aacaagttag ctttttgcaa
     1141 aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga ggccgaggcg
     1201 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
     1261 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
     1321 ttgagatgcg gatccgctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca
     1381 acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc
     1441 cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg
     1501 accatgatta cgccaagctt gggctgcagg tcgagggatc tccataagag aagagggaca
     1561 gctatgactg ggagtagtca ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa
     1621 aattttaggg atgttaaaga aaaaaataac acaaaacaaa atataaaaaa aatctaacct
     1681 caagtcaagg cttttctatg gaataaggaa tggacagcag ggggctgttt catatactga
     1741 tgacctcttt atagccaacc tttgttcatg gcagccagca tatgggcata tgttgccaaa
     1801 ctctaaacca aatactcatt ctgatgtttt aaatgatttg ccctcccata tgtccttccg
     1861 agtgagagac acaaaaaatt ccaacacact attgcaatga aaataaattt cctttattag
     1921 ccagaagtca gatgctcaag gggcttcatg atgtccccat aatttttggc agagggaaaa
     1981 agatctcagt ggtatttgtg agccagggca ttggccacac cagccaccac cttctgatag
     2041 gcagcctgca cctgaggagt gcggccggcc gcgatgtggc ttttggccgc agaggcacct
     2101 gtcgccaggt ggggggttcc gctgcctgca aagggtcgct acagacgttg tttgtcttca
     2161 agaagcttcc agaggaactg cttccttcac gacattcaac agaccttgca ttcctttggc
     2221 gagaggggaa agacccctag gaatgctcgt caagaagaca gggccaggtt tccgggccct
     2281 cacattgcca aaagacggca atatggtgga aaataacata tagacaaacg cacaccggcc
     2341 ttattccaag cggcttcggc cagtaacgtt aggggggggg gagggagagg gggggggggg
     2401 ggggcggaat tggccgcttt acttgtacag ctcgtccatg ccgtacagga acaggtggtg
     2461 gcggccctcg gagcgctcgt actgttccac gatggtgtag tcctcgttgt gggaggtgat
     2521 gtccagcttg gtgtccacgt agtagtagcc gggcagttgc acgggcttct tggccatgta
     2581 gatggtcttg aactccacca ggtagtggcc gccgtccttc agcttcaggg cctggtggat
     2641 ctcgcccttc agcacgccgt cgcgggggta caggcgctcg gtggaggcct cccagcccat
     2701 ggtcttcttc tgcattacgg ggccgtcggg ggggaagttg gtgccgcgca tcttcacctt
     2761 gtagatcagc gtgccgtcct gcagggagga gtcctgggtc acggtcacca gaccgccgtc
     2821 ctcgaagttc atcacgcgct cccacttgaa gccctcgggg aaggacagct tcttgtaatc
     2881 ggggatgtcg gcggggtgct tcacgtacgc cttggagccg tacatgaact ggggggacag
     2941 gatgtcccag gcgaagggca gggggccgcc cttggtcacc ttcagcttgg cggtctgggt
     3001 gccctcgtag gggcggccct cgccctcgcc ctcgatctcg aactcgtggc cgttcatgga
     3061 gccctccatg cgcaccttga agcgcatgaa ctctttgatg acctcctcgc ccttgctcac
     3121 catggtggcg ggatcgggcc cgatccaggc tagaaaaccg gtgcgcgcat aaagcgcctc
     3181 gcaaatcgcg tccgacgggc aactaacggc tccggtcaag cttccattat atagaattct
     3241 ttgccaaaat gatgagacag cacaataacc agcacgttgc ccaggagctg taggaaaaag
     3301 aagaaggcat gaacatggtt agcagaggct ctagagccgc cggtcacacg ccagaagccg
     3361 aaccccgccc tgccccgtcc cccccgaagg cagccgtccc cccgcggaca gccccgaggc
     3421 tggagaggga gaaggggacg gcggcgcggc gacgcacgaa ggccctcccc gcccatttcc
     3481 ttcctgccgg cgccgcaccg cttcgccccg cgcccgctag agggggtgcg gcggcgcctc
     3541 ccagattccg gctccgcaca gatttgggac aaaggaagtc cctgcgccct cccgcacgat
     3601 taccataaaa ggcaatggct gcggctcgcc gcgcctcgcc agccgccggc gctccggggg
     3661 ccgccgcgcc cctcccccga gccctccccg gcccgaggcg gccccgcccc gcccggcacc
     3721 cccacctgcc gccacccccc gcccggcacg gcgagccccg cgccacgccc cgtacggagc
     3781 cccgcacccg aagccgggcc gtgctcagca actcggggag gggggtgcag ggggggttgc
     3841 agcccgaccg acgcgcccac accccctgct caccccccca cgcacacacc ccgcacgcag
     3901 cctttgttcc cctcgcagcc ccccccgcac cgcggggcac cgcccccggc cgcgctcccc
     3961 tcgcgcacac tgcggagcgc acaaagcccc gcgccgcgcc cgcagcgctc acagccgccg
     4021 ggcagcgcgg agccgcacgc ggcgctcccc acgcacacac acacgcacgc accccccgag
     4081 ccgctccccc cgcacaaagg gccctcccgg agcccctcaa ggctttcacg cagccacaga
     4141 aaagaaacaa gccgtcatta aaccaagcgc taattacagc ccggaggaga agggccgtcc
     4201 cgcccgctca cctgtgggag taacgcggtc agtcagagcc ggggcgggcg gcgcgaggcg
     4261 gcggcggagc ggggcacggg gcgaaggcag cgcgcagcga ctcccgcccg ccgcgcgctt
     4321 cgctttttat agggccgccg ccgccgccgc ctcgccataa aaggaaactt tcggagcgcg
     4381 ccgctctgat tggctgccgc cgcacctctc cgcctcgccc cgccccgccc cccccccccc
     4441 cccccccccc cccccccccc cccccccccc cccccccccc cccccccggg cccccccccc
     4501 cccccccccc cgcccccatc gctgcacaaa ataattaaaa aataaataaa tacaaaattg
     4561 ggggtgggga ggggggggag atggggagag tgaagcagaa cgtggggctc acctcgacca
     4621 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     4681 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     4741 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     4801 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     4861 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     4921 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     4981 tagtcaataa tcaatgtcga ctatcgatag agaaatgttc tggcacctgc acttgcactg
     5041 gggacagcct attttgctag tttgttttgt ttcgttttgt tttgatggag agcgtatgtt
     5101 agtactatcg attcacacaa aaaaccaaca cacagatgta atgaaaataa agatatttta
     5161 ttgcggccgc tccaagtacc tcccgtacct taatattact tctcgaccca ggtggcactt
     5221 ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt
     5281 atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta
     5341 tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg
     5401 tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac
     5461 gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg
     5521 aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc
     5581 gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg
     5641 ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat
     5701 gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg
     5761 gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg
     5821 atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc
     5881 ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt
     5941 cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct
     6001 cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc
     6061 gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca
     6121 cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct
     6181 cactgattaa gcattggtaa ctgt
//
