LOCUS       sequence-179987-        7645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Plasmid expressing RORb2-IRES-GFPd2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7645)
  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 7645)
  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-RORb2-IRES-GFPd2
FEATURES             Location/Qualifiers
     source          1..7645
                     /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..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(3174..3748,3749..3751,3752..3760)
                     /label=IRES2
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(3341..3358)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     3568..3587
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(3761..3763,3764..3766,3767..4477,4478..4483,
                     4484..4606)
                     /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(3806..3827)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4067..4086)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4414..4435
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(4667..4686)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    4732..4787
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4786..4805)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(5148..5164)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5148..5164)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5161..5183)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5172..5188
                     /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(5196..5202,5203..5220,5221..5226))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5241..5262
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5321..5516
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      5367..5502
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    5522..5656
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5559..5578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5613..5632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5724..5741)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5895..6483)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5975..5994)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6654..7445,7446..7514))
                     /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     7277..7296
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7515..7619)
                     /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 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 attcgccacc atgtgtgaga
     1741 accagcccaa aactaaagct gacggcactg cacaaattga agtgatacca tgcaaaattt
     1801 gtggcgataa atcctccggg atccactacg gagtcatcac gtgtgaaggc tgcaagggat
     1861 tcttcaggag gagccagcag aacaatgcct cttactcctg cccaaggcag agaaactgtt
     1921 taattgacag aaccaacagg aaccgttgcc aacactgccg cctgcagaag tgtcttgccc
     1981 taggaatgtc aagagatgct gtaaagttcg ggaggatgtc caagaagcag cgggacagcc
     2041 tgtatgctga ggtgcagaag catcagcaaa ggctgcagga gcagcggcag cagcagagtg
     2101 gggaggcgga ggccctcgcc agggtgtaca gcagcagcat tagcaatggc ctcagcaacc
     2161 tgaacaccga gaccggcggc acatacgcca acgggcacgt cattgacctg cccaagtccg
     2221 aaggttatta cagcatagat tccggtcagc cgtctcccga tcagtcagga ctggacatga
     2281 ctgggatcaa acagataaag caagaaccta tctatgacct cacatctgta cccaacttgt
     2341 ttacctatag ctctttcaac aacgggcagt tagctcccgg gataacaatg tctgagatcg
     2401 atcgaattgc acagaacatc attaagtccc atttggagac atgtcagtac accatggaag
     2461 aactccatca gctggcatgg cagacccaca cctacgagga aatcaaggcg tatcaaagca
     2521 agtccaggga ggctctgtgg cagcagtgtg ccatccagat cacccatgct atccagtacg
     2581 tggtggagtt cgccaagcgg ataacaggct tcatggagct gtgtcagaac gatcagatct
     2641 tacttctgaa gtcaggttgc ttggaagtgg ttttagtgag aatgtgtcgt gccttcaacc
     2701 cattaaacaa cactgttctg tttgaaggaa aatatggagg aatgcaaatg ttcaaagcct
     2761 taggttcgga tgacctagtg aatgaagcat ttgactttgc gaagaatctg tgttccttgc
     2821 agctgactga ggaagagatt gctctgttct cctctgctgt tctgatatcc ccagaccgag
     2881 cctggctgat cgaaccaaga aaagtccaga agcttcagga aaagatttat tttgcactgc
     2941 aacatgtgat tcagaagaac cacctggatg atgagaccct ggcaaagtta atagccaaga
     3001 taccaactat cacggcagtc tgcaacttgc atggggagaa gctgcaggta tttaagcagt
     3061 ctcatccaga catagtgaat acactgtttc ctccattgta caaggagctc tttaatcctg
     3121 actgtgctgc ggtctgcaaa tgagcggccg ctacgtaaat tccgcccccc ccccccctct
     3181 ccctcccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     3241 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     3301 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     3361 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     3421 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     3481 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     3541 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     3601 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     3661 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     3721 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     3781 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     3841 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     3901 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     3961 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     4021 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     4081 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     4141 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     4201 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa
     4261 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     4321 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     4381 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     4441 cgccgggatc actctcggca tggacgagct gtacaagaag cttagccatg gcttcccgcc
     4501 ggaggtggag gagcaggatg atggcacgct gcccatgtct tgtgcccagg agagcgggat
     4561 ggaccgtcac cctgcagcct gtgcttctgc taggatcaat gtgtaggcgg ccccgccagc
     4621 acagtggtcg atcgaccaat gccctggctc acaaatacca ctgagatctt tttccctctg
     4681 ccaaaaatta tggggacatc atgaagcccc ttgagcatct gacttctggc taataaagga
     4741 aatttatttt cattgcaata gtgtgttgga attttttgtg tctctcactc ggaaggacat
     4801 atgggagggc aaatcattta aaacatcaga atgagtattt ggtttagagt ttggcaacat
     4861 atgcccatat gctggctgcc atgaacaaag gttggctata aagaggtcat cagtatatga
     4921 aacagccccc tgctgtccat tccttattcc atagaaaagc cttgacttga ggttagattt
     4981 tttttatatt ttgttttgtg ttattttttt ctttaacatc cctaaaattt tccttacatg
     5041 ttttactagc cagatttttc ctcctctcct gactactccc agtcatagct gtccctcttc
     5101 tcttatggag atccctcgac ctgcagccca agcttggcgt aatcatggtc atagctgttt
     5161 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     5221 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     5281 cccgctttcc agtcgggaaa cctgtcgtgc cagcggatcc gcatctcaat tagtcagcaa
     5341 ccatagtccc gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt
     5401 ctccgcccca tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct
     5461 ctgagctatt ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaaaagc
     5521 taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac
     5581 aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc
     5641 ttatcatgtc tggatccgct gcattaatga atcggccaac gcgcggggag aggcggtttg
     5701 cgtattgggc gctcttccgc ttcctcgctc actgactcgc tgcgctcggt cgttcggctg
     5761 cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt tatccacaga atcaggggat
     5821 aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc
     5881 gcgttgctgg cgtttttcca taggctccgc ccccctgacg agcatcacaa aaatcgacgc
     5941 tcaagtcaga ggtggcgaaa cccgacagga ctataaagat accaggcgtt tccccctgga
     6001 agctccctcg tgcgctctcc tgttccgacc ctgccgctta ccggatacct gtccgccttt
     6061 ctcccttcgg gaagcgtggc gctttctcat agctcacgct gtaggtatct cagttcggtg
     6121 taggtcgttc gctccaagct gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc
     6181 gccttatccg gtaactatcg tcttgagtcc aacccggtaa gacacgactt atcgccactg
     6241 gcagcagcca ctggtaacag gattagcaga gcgaggtatg taggcggtgc tacagagttc
     6301 ttgaagtggt ggcctaacta cggctacact agaagaacag tatttggtat ctgcgctctg
     6361 ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt gatccggcaa acaaaccacc
     6421 gctggtagcg gtggtttttt tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct
     6481 caagaagatc ctttgatctt ttctacgggg tctgacgctc agtggaacga aaactcacgt
     6541 taagggattt tggtcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     6601 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     6661 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     6721 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     6781 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     6841 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     6901 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     6961 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     7021 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     7081 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     7141 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     7201 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     7261 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     7321 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     7381 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     7441 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     7501 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     7561 ctcatgagcg gatacatatt tgaatgtatt tagaaaaata aacaaatagg ggttccgcgc
     7621 acatttcccc gaaaagtgcc acctg
//
