LOCUS       sequence-139262-        6121 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Cre-dependent reporter plasmid for bipolar cells in the retina.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6121)
  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 6121)
  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: Chx10BP-LCL-EGFP
FEATURES             Location/Qualifiers
     source          1..6121
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    46..94
                     /note="synthetic polyadenylation signal"
     misc_feature    108..199
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     148..167
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     protein_bind    complement(1043..1076)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     CDS             1077..1787
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKXKKPVQLPGAYNVNIXLDITSHNEDYTIVEQYXRA
                     EGRHSTGGMDELYK"
     primer_bind     complement(1222..1240)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     1490..1509
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    1944..1999
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     protein_bind    complement(2339..2372)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     regulatory      2396..2405
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2402..2404,2405..2407,2408..3121)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(2402..2404,2405..2407,2408..3118)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2447..2468)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2708..2727)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3055..3076
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3207..3262
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3623..3639)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3623..3639)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3636..3658)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3647..3663
                     /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(3671..3677,3678..3695,3696..3701))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3717..3738
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        3797..3992
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3843..3978
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    3998..4132
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4035..4054)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4089..4108
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4200..4217)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4371..4959)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4451..4470)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5130..5921,5922..5990))
                     /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     5753..5772
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5991..6095)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgagaagt aatattaagg tacgggaggt acttggagcg gccgcaataa aatatcttta
       61 ttttcattac atctgtgtgt tggttttttg tgtgaatcga tagtactaac atacgctctc
      121 catcaaaaca aaacgaaaca aaacaaacta gcaaaatagg ctgtccccag tgcaagtgca
      181 ggtgccagaa catttctcta tcgatagtcg accaattcga ttcacactca gttcttcagg
      241 gaagagtcaa tcccggcctt ctcctcactc ccccccaccc ggcagaacct catctccatc
      301 tgcttgatca gggcctgcac tgtatattgc agcaatttta aattgcacca ttaaagattt
      361 tatgctggag acgttaaaat taaaaagatc tatgtgcaaa gagtcacaac gaatttaatc
      421 aacttgattt aatagaaaac agcaaatgga atttatgatg ctccaaaaga caagggaatt
      481 tttttcccac agaaaaataa taaaaaagca ctttgtagaa aacagaaagt ggagtacggg
      541 ggggggggct ccccatctga gagtttatca gaacctaatc cggggagaag agcactggct
      601 ggggctgctt gcccgctaat cccagctgcc attaaaatat taaagataaa tctaatcgtc
      661 tctttatcca aaataagcga cttttgtgtg gggagaaaac gtctaacccc ttaggaggag
      721 aattagttct aatgcatcaa atggaattcg gtccaggctg aggcgaaata gggtttagag
      781 gcaaatgaga cggtaataga actaaaaccc agaatgacaa attaaaaacc ctcatttaca
      841 cgaattaaaa cggctcacaa aagggataga aacccagctc tgccctcctc agcccccaac
      901 ctgccttcct gtcctataat cactagtgaa ttgctcgagg aattctatat aatggaagct
      961 tgaccggagc cgttagttgc ccgtcggacg cgatttgcga ggcgctttat gcgcgcaccg
     1021 gtacgactca ctataggcta gcataacttc gtatagcata cattatacga agttatatgg
     1081 tgagcaaggg cgaggaggat aacatggcca tcatcaagga gttcatgcgc ttcaaggtgc
     1141 acatggaggg ctccgtgaac ggccacgagt tcgagatcga gggcgagggc gagggccgcc
     1201 cctacgaggg cacccagacc gccaagctga aggtgaccaa gggtggcccc ctgcccttcg
     1261 cctgggacat cctgtcccct cagttcatgt acggctccaa ggcctacgtg aagcaccccg
     1321 ccgacatccc cgactacttg aagctgtcct tccccgaggg cttcaagtgg gagcgcgtga
     1381 tgaacttcga ggacggcggc gtggtgaccg tgacccagga ctcctccctg caggacggcg
     1441 agttcatcta caaggtgaag ctgcgcggca ccaacttccc ctccgacggc cccgtaatgc
     1501 agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag gacggcgccc
     1561 tgaagggcga gatcaagcag aggctgaagc tgaaggacgg cggccactac gacgctgagg
     1621 tcaagaccac ctacaaggnc aagaagcccg tgcagctgcc cggcgcctac aacgtcaaca
     1681 tcnagttgga catcacctcc cacaacgagg actacaccat cgtggaacag tacnaacgcg
     1741 ccgagggccg ccactccacc ggcggcatgg acgagctgta caagtaaaca cgtgcggccg
     1801 cactcctcag gtgcaggctg cctatcagaa ggtggtggct ggtgtggcca atgccctggc
     1861 tcacaaatac cactgagatc tttttccctc tgccaaaaat tatggggaca tcatgaagcc
     1921 ccttgagcat ctgacttctg gctaataaag gaaatttatt ttcattgcaa tagtgtgttg
     1981 gaattttttg tgtctctcac tcggaaggac atatgggagg gcaaatcatt taaaacatca
     2041 gaatgagtat ttggtttaga gtttggcaac atatgcccat atgctggctg ccatgaacaa
     2101 aggttggcta taaagaggtc atcagtatat gaaacagccc cctgctgtcc attccttatt
     2161 ccatagaaaa gccttgactt gaggttagat tttttttata ttttgttttg tgttattttt
     2221 ttctttaaca tccctaaaat tttccttaca tgttttacta gccagatttt tcctcctctc
     2281 ctgactactc ccagtcatag ctgtccctct tctcttatgg agatccctcg acctgcagat
     2341 aacttcgtat agcatacatt atacgaagtt atactagcct cgagaattca cgcgggccgc
     2401 catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg tcgagctgga
     2461 cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg atgccaccta
     2521 cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc cctggcccac
     2581 cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg accacatgaa
     2641 gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc gcaccatctt
     2701 cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg gcgacaccct
     2761 ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca tcctggggca
     2821 caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca agcagaagaa
     2881 cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg tgcagctcgc
     2941 cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc ccgacaacca
     3001 ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg atcacatggt
     3061 cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc tgtacaagta
     3121 ataagctcgc gtggtacctg atctttttcc ctctgccaaa aattatgggg acatcatgaa
     3181 gccccttgag catctgactt ctggctaata aaggaaattt attttcattg caatagtgtg
     3241 ttggaatttt ttgtgtctct cactcggaag gacatatggg agggcaaatc atttaaaaca
     3301 tcagaatgag tatttggttt agagtttggc aacatatgcc catatgctgg ctgccatgaa
     3361 caaaggttgg ctataaagag gtcatcagta tatgaaacag ccccctgctg tccattcctt
     3421 attccataga aaagccttga cttgaggtta gatttttttt atattttgtt ttgtgttatt
     3481 tttttcttta acatccctaa aattttcctt acatgtttta ctagccagat ttttcctcct
     3541 ctcctgacta ctcccagtca tagctgtccc tcttctctta tggagatccc tcgacctgca
     3601 gcccaagctt ggcgtaatca tggtcatagc tgtttcctgt gtgaaattgt tatccgctca
     3661 caattccaca caacatacga gccggaagca taaagtgtaa agcctggggg tgcctaatga
     3721 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     3781 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3841 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3901 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     3961 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     4021 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4081 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     4141 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     4201 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     4261 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     4321 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     4381 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     4441 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     4501 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     4561 tctcaatgct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     4621 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     4681 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4741 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     4801 tacactagaa ggacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4861 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4921 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4981 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     5041 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     5101 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     5161 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     5221 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     5281 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     5341 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     5401 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     5461 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     5521 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     5581 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     5641 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     5701 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     5761 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     5821 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     5881 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     5941 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     6001 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     6061 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     6121 g
//
