LOCUS       sequence-179982-        6731 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  B108 enhancer of Blimp1 gene drives EGFP expression..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6731)
  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 6731)
  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: B108-GFP-IRES-AP
FEATURES             Location/Qualifiers
     source          1..6731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     polyA_signal    47..95
                     /note="synthetic polyadenylation signal"
     misc_feature    109..200
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     149..168
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     regulatory      420..429
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(426..428,429..431,432..1145)
                     /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(426..428,429..431,432..1142)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(471..492)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(732..751)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1079..1100
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    1172..1745
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    1195..1747
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1339..1356)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1566..1585
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             1759..3258
                     /codon_start=1
                     /product="secreted alkaline phosphatase from human 
                     placenta"
                     /label=SEAP
                     /note="The natural enzyme has a C-terminal transmembrane 
                     sequence that is replaced by a GPI anchor."
                     /translation="LLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQPA
                     QTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVDKH
                     VPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGVVT
                     TTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGRKY
                     MFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVTHL
                     MGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHESRA
                     YRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGKAR
                     DRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVFAR
                     GPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD"
     primer_bind     complement(3753..3772)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    3818..3873
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(3872..3891)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(4234..4250)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4234..4250)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4247..4269)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4258..4274
                     /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(4282..4288,4289..4306,4307..4312))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4327..4348
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        4407..4602
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4453..4588
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    4608..4742
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4645..4664)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4699..4718
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4810..4827)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4981..5569)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5061..5080)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5740..6531,6532..6600))
                     /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     6363..6382
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6601..6705)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ggtcgagaag taatattaag gtacgggagg tacttggagc ggccgcaata aaatatcttt
       61 attttcatta catctgtgtg ttggtttttt gtgtgaatcg atagtactaa catacgctct
      121 ccatcaaaac aaaacgaaac aaaacaaact agcaaaatag gctgtcccca gtgcaagtgc
      181 aggtgccaga acatttctct atcgatagtc gaccaattgc tcgagttcgg aaataggtca
      241 ggtacttggg agattagaca agcttgcttc atccctgaat acaattagga ggaattagac
      301 ctgcagcaag tagcttggat ttatcagctt taattgaatt ctatataatg gaagcttgac
      361 cggagccgtt agttgcccgt cggacgcgat ttgcgaggcg ctttatgcgc gcaccggtcg
      421 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
      481 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
      541 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
      601 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
      661 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
      721 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
      781 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
      841 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
      901 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
      961 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1021 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca
     1081 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1141 agtaaagcgg ccaattccgc cccccccccc ccccctctcc ctcccccccc cctaacgtta
     1201 ctggccgaag ccgcttggaa taaggccggt gtgcgtttgt ctatatgtta ttttccacca
     1261 tattgccgtc ttttggcaat gtgagggccc ggaaacctgg ccctgtcttc ttgacgagca
     1321 ttcctagggg tctttcccct ctcgccaaag gaatgcaagg tctgttgaat gtcgtgaagg
     1381 aagcagttcc tctggaagct tcttgaagac aaacaacgtc tgtagcgacc ctttgcaggc
     1441 agcggaaccc cccacctggc gacaggtgcc tctgcggcca aaagccacgt gtataagata
     1501 cacctgcaaa ggcggcacaa ccccagtgcc acgttgtgag ttggatagtt gtggaaagag
     1561 tcaaatggct ctcctcaagc gtattcaaca aggggctgaa ggatgcccag aaggtacccc
     1621 attgtatggg atctgatctg gggcctcggt gcacatgctt tacatgtgtt tagtcgaggt
     1681 taaaaaaacg tctaggcccc ccgaaccacg gggacgtggt tttcctttga aaaacacgat
     1741 gataatatgg ggtcgaggct gctgctgctg ctgctgctgg gcctgaggct acagctctcc
     1801 ctgggcatca tcccagttga ggaggagaac ccggacttct ggaaccgcga ggcagccgag
     1861 gccctgggtg ccgccaagaa gctgcagcct gcacagacag ccgccaagaa cctcatcatc
     1921 ttcctgggcg atgggatggg ggtgtctacg gtgacagctg ccaggatcct aaaagggcag
     1981 aagaaggaca aactggggcc tgagataccc ctggccatgg accgcttccc atatgtggct
     2041 ctgtccaaga catacaatgt agacaaacat gtgccagaca gtggagccac agccacggcc
     2101 tacctgtgcg gggtcaaggg caacttccag accattggct tgagtgcagc cgcccgcttt
     2161 aaccagtgca acacgacacg cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa
     2221 gcagggaagt cagtgggagt ggtaaccacc acacgagtgc agcacgcctc gccagccggc
     2281 acctacgccc acacggtgaa ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc
     2341 cgccaggagg ggtgccagga catcgctacg cagctcatct ccaacatgga cattgacgtg
     2401 atcctaggtg gaggccgaaa gtacatgttt cgcatgggaa ccccagaccc tgagtaccca
     2461 gatgactaca gccaaggtgg gaccaggctg gacgggaaga atctggtgca ggaatggctg
     2521 gcgaagcgcc agggtgcccg gtatgtgtgg aaccgcactg agctcatgca ggcttccctg
     2581 gacccgtctg tgacccatct catgggtctc tttgagcctg gagacatgaa atacgagatc
     2641 caccgagact ccacactgga cccctccctg atggagatga cagaggctgc cctgcgcctg
     2701 ctgagcagga acccccgcgg cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt
     2761 catcatgaaa gcagggctta ccgggcactg actgagacga tcatgttcga cgacgccatt
     2821 gagagggcgg gccagctcac cagcgaggag gacacgctga gcctcgtcac tgccgaccac
     2881 tcccacgtct tctccttcgg aggctacccc ctgcgaggga gctccatctt cgggctggcc
     2941 cctggcaagg cccgggacag gaaggcctac acggtcctcc tatacggaaa cggtccaggc
     3001 tatgtgctca aggacggcgc ccggccggat gttaccgaga gcgagagcgg gagccccgag
     3061 tatcggcagc agtcagcagt gcccctggac gaagagaccc acgcaggcga ggacgtggcg
     3121 gtgttcgcgc gcggcccgca ggcgcacctg gttcacggcg tgcaggagca gaccttcata
     3181 gcgcacgtca tggccttcgc cgcctgcctg gagccctaca ccgcctgcga cctggcgccc
     3241 cccgccggca ccaccgacgc cgcgcacccg gggcggtccg tggtccccgc gttgcttcct
     3301 ctgctggccg ggaccctgct gctgctggag acggccactg ctccctgagt gtcccgtccc
     3361 tggggctcct gcttccccat cccggagttc tcctgctccc cacctcctgt cgtcctgcct
     3421 ggcctccagc ccgagtcgtc atccccggag tccctataca gaggtcctgc catggaacct
     3481 tcccctcccc gtgcgctctg gggactgagc ccatgacacc aaacctgccc cttggctgct
     3541 ctcggactcc ctaccccaac cccagggact gcaggttgtg ccctgtggct gcctgcaccc
     3601 caggaaagga gggggctcag gccatccagc caccacctac agcccagtgg ggtcgaggat
     3661 atcgcggccg gccgcactcc tcaggtgcag gctgcctatc agaaggtggt ggctggtgtg
     3721 gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg
     3781 gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt
     3841 gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat
     3901 catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg
     3961 gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct
     4021 gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt
     4081 tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga
     4141 tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc
     4201 ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg
     4261 ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg
     4321 tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc
     4381 gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc
     4441 ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc
     4501 tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag
     4561 aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg
     4621 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4681 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     4741 tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     4801 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     4861 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     4921 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     4981 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     5041 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     5101 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     5161 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     5221 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     5281 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     5341 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     5401 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     5461 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg
     5521 tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt
     5581 gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt
     5641 catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa
     5701 atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga
     5761 ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt
     5821 gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg
     5881 agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga
     5941 gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga
     6001 agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg
     6061 catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc
     6121 aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc
     6181 gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca
     6241 taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac
     6301 caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg
     6361 ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc
     6421 ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg
     6481 tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac
     6541 aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat
     6601 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
     6661 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
     6721 agtgccacct g
//
