LOCUS       sequence-179986-        4849 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid expressing RORb2-IRES-GFPd2.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4849)
  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 4849)
  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..4849
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     misc_feature    78..540
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(134..151)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     361..380
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(554..556,557..559,560..1270,1271..1276,1277..1399)
                     /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(599..620)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(860..879)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1207..1228
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1460..1479)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1525..1580
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1579..1598)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1941..1957)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1941..1957)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1954..1976)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1965..1981
                     /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(1989..1995,1996..2013,2014..2019))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2034..2055
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2114..2309
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2160..2295
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2315..2449
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2352..2371)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2406..2425
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2517..2534)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2688..3276)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2768..2787)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3447..4238,4239..4307))
                     /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     4070..4089
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4308..4412)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        4443..4822
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 cgaagccgct tggaataagg ccggtgtgcg tttgtctata tgttattttc caccatattg
       61 ccgtcttttg gcaatgtgag ggcccggaaa cctggccctg tcttcttgac gagcattcct
      121 aggggtcttt cccctctcgc caaaggaatg caaggtctgt tgaatgtcgt gaaggaagca
      181 gttcctctgg aagcttcttg aagacaaaca acgtctgtag cgaccctttg caggcagcgg
      241 aaccccccac ctggcgacag gtgcctctgc ggccaaaagc cacgtgtata agatacacct
      301 gcaaaggcgg cacaacccca gtgccacgtt gtgagttgga tagttgtgga aagagtcaaa
      361 tggctctcct caagcgtatt caacaagggg ctgaaggatg cccagaaggt accccattgt
      421 atgggatctg atctggggcc tcggtgcaca tgctttacat gtgtttagtc gaggttaaaa
      481 aaacgtctag gccccccgaa ccacggggac gtggttttcc tttgaaaaac acgatgataa
      541 tatggccaca accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct
      601 ggtcgagctg gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg
      661 cgatgccacc tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt
      721 gccctggccc accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc
      781 cgaccacatg aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga
      841 gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga
      901 gggcgacacc ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa
      961 catcctgggg cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga
     1021 caagcagaag aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag
     1081 cgtgcagctc gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct
     1141 gcccgacaac cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg
     1201 cgatcacatg gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga
     1261 gctgtacaag aagcttagcc atggcttccc gccggaggtg gaggagcagg atgatggcac
     1321 gctgcccatg tcttgtgccc aggagagcgg gatggaccgt caccctgcag cctgtgcttc
     1381 tgctaggatc aatgtgtagg cggccccgcc agcacagtgg tcgatcgacc aatgccctgg
     1441 ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc
     1501 cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt
     1561 ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc
     1621 agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca
     1681 aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat
     1741 tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt
     1801 tttctttaac atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct
     1861 cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc
     1921 ccaagcttgg cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca
     1981 attccacaca acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg
     2041 agctaactca cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg
     2101 tgccagcgga tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     2161 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     2221 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     2281 gcttttttgg aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg
     2341 ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc
     2401 tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa
     2461 tgaatcggcc aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg
     2521 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     2581 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     2641 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     2701 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     2761 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     2821 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     2881 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     2941 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     3001 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     3061 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     3121 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     3181 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     3241 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     3301 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     3361 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     3421 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     3481 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     3541 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     3601 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     3661 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     3721 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     3781 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     3841 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     3901 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     3961 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     4021 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     4081 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     4141 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     4201 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     4261 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     4321 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     4381 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctggg
     4441 tcgacattga ttattgacta gttattaata gtaatcaatt acggggtcat tagttcatag
     4501 cccatatatg gagttccgcg ttacataact tacggtaaat ggcccgcctg gctgaccgcc
     4561 caacgacccc cgcccattga cgtcaataat gacgtatgtt cccatagtaa cgccaatagg
     4621 gactttccat tgacgtcaat gggtggagta tttacggtaa actgcccact tggcagtaca
     4681 tcaagtgtat catatgccaa gtacgccccc tattgacgtc aatgacggta aatggcccgc
     4741 ctggcattat gcccagtaca tgaccttatg ggactttcct acttggcagt acatctacgt
     4801 attagtcatc gctattacca tggtcgaggt gagccccacg ttctgcttc
//
