LOCUS       sequence-179993-        7125 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing Crx-IRES- GFPd2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7125)
  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 7125)
  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-Crx-IRES-GFPd2
FEATURES             Location/Qualifiers
     source          1..7125
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..383
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        386..661
                     /label=chicken beta-actin promoter
     intron          662..1670
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and 
                     rabbit beta-globin"
     primer_bind     1594..1616
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     1678..1697
                     /label=pCAG-F
                     /note="Rabbit beta-globin intron, for pCAG plasmids, 
                     forward primer"
     misc_feature    join(2654..3228,3229..3231,3232..3240)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(2821..2838)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3048..3067
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3241..3243,3244..3246,3247..3957,3958..3963,
                     3964..4086)
                     /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(3286..3307)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3547..3566)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3894..3915
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4147..4166)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4212..4267
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4266..4285)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4628..4644)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4628..4644)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4641..4663)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4652..4668
                     /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(4676..4682,4683..4700,4701..4706))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4721..4742
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4801..4996
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4847..4982
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5002..5136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5039..5058)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5093..5112
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5204..5221)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5375..5963)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5455..5474)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6134..6925,6926..6994))
                     /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     6757..6776
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6995..7099)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agttattaat agtaatcaat tacggggtca ttagttcata
       61 gcccatatat ggagttccgc gttacataac ttacggtaaa tggcccgcct ggctgaccgc
      121 ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt tcccatagta acgccaatag
      181 ggactttcca ttgacgtcaa tgggtggact atttacggta aactgcccac ttggcagtac
      241 atcaagtgta tcatatgcca agtacgcccc ctattgacgt caatgacggt aaatggcccg
      301 cctggcatta tgcccagtac atgaccttat gggactttcc tacttggcag tacatctacg
      361 tattagtcat cgctattacc atgggtcgag 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 tggcggagcc gaaatctggg aggcgccgcc gcaccccctc
     1441 tagcgggcgc gggcgaagcg gtgcggcgcc ggcaggaagg aaatgggcgg ggagggcctt
     1501 cgtgcgtcgc cgcgccgccg tccccttctc catctccagc ctcggggctg ccgcaggggg
     1561 acggctgcct tcggggggga cggggcaggg cggggttcgg cttctggcgt gtgaccggcg
     1621 gctctagagc ctctgctaac catgttcatg ccttcttctt tttcctacag ctcctgggca
     1681 acgtgctggt tattgtgctg tctcatcatt ttggcaaaga attgatgatg gcatatatga
     1741 acccggggcc tcactattca gtcaatgcct tggctctgag tggccccaat gtggacctga
     1801 tgcaccaggc tgtcccatac tcaagtgccc ctaggaagca gcggcgggag cggaccacat
     1861 tcaccaggag ccagctggag gagctggagg ccctgtttgc caagacccag tacccggatg
     1921 tgtatgcacg tgaggaggtt gctcttaaga tcaatctgcc tgagtccagg gtccaggtct
     1981 ggttcaagaa tcgtagggcg aaatgcagac agcagcgaca gcagcagaaa cagcaacagc
     2041 agcccccggg ggcacagacc aaggctcgtc ctgcgaagag gaaggcaggg acatccccga
     2101 gaccctctac agatgtttgt acagatcctt tgggcatctc agattcttac agcccatctc
     2161 tgcctggccc ctcaggttca cctaccacag cagtggccac cgtgtccatt tggagtccag
     2221 cctcagaggc tcctttgcct gaggcccaga gagccggcct agtggcttct gggccctctc
     2281 tcacctcagc cccttatgcc atgacctatg ccccggcttc tgctttctgt tcttcccctt
     2341 cagcttatgc atctccaagt tcctatttca gtgggttgga tccctacctt tcgcccatgg
     2401 tgccccaact tgggggtccg gctctcagcc ccctctcagg cccctctgtg gggccatccc
     2461 tggcccagtc ccccacctcc ttgtcaggcc agagctatag cacctacagt cctgtggaca
     2521 gcctggaatt caaggacccc acaggcacct ggaaatttac ctacaatccc atggaccctc
     2581 tggactacaa agatcagagt gcctggaagt ttcagatctt gtagcggccg ctacgtaaat
     2641 tccgcccccc ccccccctct ccctcccccc cccctaacgt tactggccga agccgcttgg
     2701 aataaggccg gtgtgcgttt gtctatatgt tattttccac catattgccg tcttttggca
     2761 atgtgagggc ccggaaacct ggccctgtct tcttgacgag cattcctagg ggtctttccc
     2821 ctctcgccaa aggaatgcaa ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag
     2881 cttcttgaag acaaacaacg tctgtagcga ccctttgcag gcagcggaac cccccacctg
     2941 gcgacaggtg cctctgcggc caaaagccac gtgtataaga tacacctgca aaggcggcac
     3001 aaccccagtg ccacgttgtg agttggatag ttgtggaaag agtcaaatgg ctctcctcaa
     3061 gcgtattcaa caaggggctg aaggatgccc agaaggtacc ccattgtatg ggatctgatc
     3121 tggggcctcg gtgcacatgc tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc
     3181 ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat ggccacaacc
     3241 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     3301 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     3361 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     3421 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     3481 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     3541 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     3601 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     3661 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     3721 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     3781 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     3841 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     3901 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagaag
     3961 cttagccatg gcttcccgcc ggaggtggag gagcaggatg atggcacgct gcccatgtct
     4021 tgtgcccagg agagcgggat ggaccgtcac cctgcagcct gtgcttctgc taggatcaat
     4081 gtgtaggcgg ccccgccagc acagtggtcg atcgaccaat gccctggctc acaaatacca
     4141 ctgagatctt tttccctctg ccaaaaatta tggggacatc atgaagcccc ttgagcatct
     4201 gacttctggc taataaagga aatttatttt cattgcaata gtgtgttgga attttttgtg
     4261 tctctcactc ggaaggacat atgggagggc aaatcattta aaacatcaga atgagtattt
     4321 ggtttagagt ttggcaacat atgcccatat gctggctgcc atgaacaaag gttggctata
     4381 aagaggtcat cagtatatga aacagccccc tgctgtccat tccttattcc atagaaaagc
     4441 cttgacttga ggttagattt tttttatatt ttgttttgtg ttattttttt ctttaacatc
     4501 cctaaaattt tccttacatg ttttactagc cagatttttc ctcctctcct gactactccc
     4561 agtcatagct gtccctcttc tcttatggag atccctcgac ctgcagccca agcttggcgt
     4621 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4681 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4741 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc
     4801 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     4861 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     4921 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     4981 cctaggcttt tgcaaaaagc taacttgttt attgcagctt ataatggtta caaataaagc
     5041 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5101 tccaaactca tcaatgtatc ttatcatgtc tggatccgct gcattaatga atcggccaac
     5161 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     5221 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     5281 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     5341 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     5401 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     5461 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     5521 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     5581 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     5641 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     5701 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     5761 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     5821 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     5881 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     5941 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     6001 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     6061 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     6121 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     6181 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     6241 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     6301 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     6361 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     6421 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     6481 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     6541 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     6601 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     6661 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     6721 ggcgaccgag ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa
     6781 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     6841 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     6901 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     6961 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     7021 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     7081 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctg
//
