LOCUS       sequence-179994-        7099 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Otx2(isoform b)-IRES-GFPd2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7099)
  AUTHORS   Wang S, Sengel C, Emerson MM, Cepko CL
  TITLE     A gene regulatory network controls the binary fate decision of rod 
            and bipolar cells in the vertebrate retina.
  JOURNAL   Dev Cell. 2014 Sep 8;30(5):513-27. doi: 
            10.1016/j.devcel.2014.07.018. Epub 2014 Aug 21.
  PUBMED    25155555
REFERENCE   2  (bases 1 to 7099)
  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: 
            "10.1016/j.devcel.2014.07.018"; journalName: "Dev Cell"; date: 
            "2014-09-8- 8"; volume: "30"; issue: "5"; pages: "513-27"
COMMENT     Sequence Label: CAG-Otx2-IRES-GFPd2
FEATURES             Location/Qualifiers
     source          1..7099
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        5..384
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1673
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1597..1619
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1681..1700
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(2628..3202,3203..3205,3206..3214)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2795..2812)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3022..3041
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3215..3217,3218..3220,3221..3931,3932..3937,
                     3938..4060)
                     /codon_start=1
                     /product="EGFP destabilized by fusion to residues 422-461 
                     of mouse ornithine decarboxylase, giving an in vivo 
                     half-life of ~2 hours"
                     /label=d2EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYKKLSHGFPPEVEEQDDGTLPMSCAQESGMDRHPAACASARINV
                     "
     primer_bind     complement(3260..3281)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3521..3540)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3868..3889
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4121..4140)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4186..4241
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4240..4259)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4602..4618)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4602..4618)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4615..4637)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4626..4642
                     /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(4650..4656,4657..4674,4675..4680))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4695..4716
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4775..4970
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4821..4956
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4976..5110
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5013..5032)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5067..5086
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5178..5195)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5349..5937)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5429..5448)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6108..6899,6900..6968))
                     /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     6731..6750
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6969..7073)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat
       61 agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg
      121 cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata
      181 gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta
      241 catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc
      301 gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca gtacatctac
      361 gtattagtca tcgctattac catggtcgag gtgagcccca cgttctgctt cactctcccc
      421 atctcccccc cctccccacc cccaattttg tatttattta ttttttaatt attttgtgca
      481 gcgatggggg cggggggggg gggggcgcgc gccaggcggg gcggggcggg gcgaggggcg
      541 gggcggggcg aggcggagag gtgcggcggc agccaatcag agcggcgcgc tccgaaagtt
      601 tccttttatg gcgaggcggc ggcggcggcg gccctataaa aagcgaagcg cgcggcgggc
      661 gggagtcgct gcgttgcctt cgccccgtgc cccgctccgc gccgcctcgc gccgcccgcc
      721 ccggctctga ctgaccgcgt tactcccaca ggtgagcggg cgggacggcc cttctcctcc
      781 gggctgtaat tagcgcttgg tttaatgacg gctcgtttct tttctgtggc tgcgtgaaag
      841 ccttaaaggg ctccgggagg gccctttgtg cgggggggag cggctcgggg ggtgcgtgcg
      901 tgtgtgtgtg cgtggggagc gccgcgtgcg gcccgcgctg cccggcggct gtgagcgctg
      961 cgggcgcggc gcggggcttt gtgcgctccg cgtgtgcgcg aggggagcgc ggccgggggc
     1021 ggtgccccgc ggtgcggggg ggctgcgagg ggaacaaagg ctgcgtgcgg ggtgtgtgcg
     1081 tgggggggtg agcagggggt gtgggcgcgg cggtcgggct gtaacccccc cctgcacccc
     1141 cctccccgag ttgctgagca cggcccggct tcgggtgcgg ggctccgtgc ggggcgtggc
     1201 gcggggctcg ccgtgccggg cggggggtgg cggcaggtgg gggtgccggg cggggcgggg
     1261 ccgcctcggg ccggggaggg ctcgggggag gggcgcggcg gccccggagc gccggcggct
     1321 gtcgaggcgc ggcgagccgc agccattgcc ttttatggta atcgtgcgag agggcgcagg
     1381 gacttccttt gtcccaaatc tgtgcggagc cgaaatctgg gaggcgccgc cgcaccccct
     1441 ctagcgggcg cggggcgaag cggtgcggcg ccggcaggaa ggaaatgggc ggggagggcc
     1501 ttcgtgcgtc gccgcgccgc cgtccccttc tccctctcca gcctcggggc tgtccgcggg
     1561 gggacggctg ccttcggggg ggacggggca gggcggggtt cggcttctgg cgtgtgaccg
     1621 gcggctctag agcctctgct aaccatgttc atgccttctt ctttttccta cagctcctgg
     1681 gcaacgtgct ggttattgtg ctgtctcatc attttggcaa agaattgatg atgtcttatc
     1741 taaagcaacc gccttacgca gtcaatgggc tgagtctgac cacttcgggt atggacttgc
     1801 tgcatccctc cgtgggctac cccgccaccc cccggaaaca gcgaagggag aggacgacat
     1861 ttactagggc acagctcgac gttctggaag ctctgtttgc caagacccgg tacccagaca
     1921 tcttcatgag ggaagaggtg gcactgaaaa tcaacttgcc agaatccagg gtgcaggtat
     1981 ggtttaagaa tcgaagagct aagtgccgcc aacagcagca gcagcagcag aatggaggtc
     2041 agaacaaagt gaggcctgcc aagaagaaga gctctccagc tcgggaagtg agttcagaga
     2101 gtggaacaag tggccagttc agtcccccct ctagtacctc agtcccaacc attgccagca
     2161 gcagtgctcc agtgtctatc tggagcccag cgtccatctc cccactgtct gaccccttgt
     2221 ccacttcctc ctcctgcatg cagaggtcct atcccatgac ctatactcag gcttcaggtt
     2281 atagtcaagg ctatgctggc tcaacttcct actttggggg catggactgt ggatcttatt
     2341 tgacccctat gcatcaccag cttcctggac caggggccac actcagtccc atgggtacca
     2401 atgctgttac cagccatctc aatcagtccc cagcttctct ttccacccag ggatatggag
     2461 cttcaagctt gggttttaac tcaaccactg attgcttgga ttataaggac caaactgcct
     2521 cttggaagct taacttcaat gctgactgct tggattataa agatcagacg tcctcatgga
     2581 aattccaggt tttgtgagcg gccgctacgt aaattccgcc cccccccccc ctctccctcc
     2641 ccccccccta acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat
     2701 atgttatttt ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct
     2761 gtcttcttga cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg
     2821 ttgaatgtcg tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta
     2881 gcgacccttt gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag
     2941 ccacgtgtat aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg
     3001 atagttgtgg aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat
     3061 gcccagaagg taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca
     3121 tgtgtttagt cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc
     3181 ctttgaaaaa cacgatgata atatggccac aaccatggtg agcaagggcg aggagctgtt
     3241 caccggggtg gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag
     3301 cgtgtccggc gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg
     3361 caccaccggc aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt
     3421 gcagtgcttc agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat
     3481 gcccgaaggc tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac
     3541 ccgcgccgag gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat
     3601 cgacttcaag gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca
     3661 caacgtctat atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg
     3721 ccacaacatc gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat
     3781 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag
     3841 caaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg
     3901 gatcactctc ggcatggacg agctgtacaa gaagcttagc catggcttcc cgccggaggt
     3961 ggaggagcag gatgatggca cgctgcccat gtcttgtgcc caggagagcg ggatggaccg
     4021 tcaccctgca gcctgtgctt ctgctaggat caatgtgtag gcggccccgc cagcacagtg
     4081 gtcgatcgac caatgccctg gctcacaaat accactgaga tctttttccc tctgccaaaa
     4141 attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa aggaaattta
     4201 ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg acatatggga
     4261 gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca acatatgccc
     4321 atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat atgaaacagc
     4381 cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag atttttttta
     4441 tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta catgttttac
     4501 tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct cttctcttat
     4561 ggagatccct cgacctgcag cccaagcttg gcgtaatcat ggtcatagct gtttcctgtg
     4621 tgaaattgtt atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa
     4681 gcctggggtg cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct
     4741 ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca gcaaccatag
     4801 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     4861 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg gcctctgagc
     4921 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctaactt
     4981 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     5041 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     5101 tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg tttgcgtatt
     5161 gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     5221 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     5281 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     5341 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     5401 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     5461 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     5521 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     5581 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     5641 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     5701 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     5761 tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     5821 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     5881 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     5941 gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc acgttaaggg
     6001 attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga
     6061 agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta ccaatgctta
     6121 atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt tgcctgactc
     6181 cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag tgctgcaatg
     6241 ataccgcgag acccacgctc accggctcca gatttatcag caataaacca gccagccgga
     6301 agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc tattaattgt
     6361 tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt
     6421 gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag ctccggttcc
     6481 caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt tagctccttc
     6541 ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat ggttatggca
     6601 gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt gactggtgag
     6661 tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc ttgcccggcg
     6721 tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat cattggaaaa
     6781 cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa
     6841 cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt ttctgggtga
     6901 gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg gaaatgttga
     6961 atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta ttgtctcatg
     7021 agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc gcgcacattt
     7081 ccccgaaaag tgccacctg
//
