LOCUS       sequence-446856-        7613 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Enhancer reporter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7613)
  AUTHORS   Honnell V, Sweeney S, Norrie J, Parks M, Ramirez C, Jannu AJ, Xu B, 
            Teubner B, Lee AY, Bell C, Dyer MA
  TITLE     Evolutionary conservation of VSX2 super-enhancer modules in retinal 
            development.
  JOURNAL   Development. 2024 Jul 1;151(13):dev202435. doi: 10.1242/dev.202435. 
            Epub 2024 Jul 12.
  PUBMED    38994775
REFERENCE   2  (bases 1 to 7613)
  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"; journalName: 
            "Development."; date: "2024-07-1"; volume: "151(13):dev202435. doi";
            pages: " 10.1242/dev.202435. Epub 2024 Jul 12"
COMMENT     Sequence Label: Vsx2 hR0-36 +SV40 base prom-GFP-IRES-AP
FEATURES             Location/Qualifiers
     source          1..7613
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        45..240
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      91..226
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    246..380
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(283..302)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     337..356
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(448..465)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(619..1207)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(699..718)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(1378..2169,2170..2238))
                     /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     2001..2020
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2239..2343)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3441..3637
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3488..3623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      3672..3681
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3678..3680,3681..3683,3684..4397)
                     /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(3678..3680,3681..3683,3684..4394)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3723..3744)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3984..4003)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4331..4352
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    4423..4996
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    4446..4998
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(4590..4607)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     4817..4836
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             5010..6509
                     /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(7004..7023)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7069..7124
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7123..7142)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(7485..7501)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7485..7501)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(7498..7520)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    7509..7525
                     /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(7533..7539,7540..7557,7558..7563))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7578..7599
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 actgcccgct ttccagtcgg gaaacctgtc gtgccagcgg atccgcatct caattagtca
       61 gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc
      121 cattctccgc cccatggctg actaattttt tttatttatg cagaggccga ggccgcctcg
      181 gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa
      241 aagctaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
      301 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
      361 tatcttatca tgtctggatc cgctgcatta atgaatcggc caacgcgcgg ggagaggcgg
      421 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
      481 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
      541 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
      601 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
      661 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
      721 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
      781 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
      841 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
      901 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
      961 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     1021 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     1081 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     1141 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     1201 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     1261 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     1321 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     1381 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     1441 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     1501 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     1561 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     1621 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     1681 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     1741 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     1801 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     1861 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     1921 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     1981 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     2041 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     2101 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     2161 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     2221 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     2281 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     2341 gcgcacattt ccccgaaaag tgccacctgg gtcgacatgg actggaactg atttttccta
     2401 tctatgtagg ctttttggca tttggggtgt gtgtccttgt ctatgcctaa agaggccact
     2461 ggccttccca ccctttgcct ggcgcttttt cctctcgagc agctgggggc ttcagacgat
     2521 accaggtgta actgtgaggc tcctggttgg cgtgagtgct ggcaaagcct gttaattagc
     2581 acagaaacac cccagtgctc cattagctgg gccaggcatt tggaaccttc ccctcctgtg
     2641 ggccccttcc tcctaatgag agctggaaga agggccctca caaaacactg cctcagaata
     2701 atattctgct ttgtccccac cagtaaccag ggctgacagg atcactctgt ccatttctct
     2761 tctaattagc acaattccag catggcctga gccagcaaat agagaaaagc gatcaaaagc
     2821 gatcggacaa ccgtaccgaa tcacaaacac aaagagggcc attccctgta attaggtgag
     2881 acaattcctc agtcttttct cggctggagt ttgccgctaa gactcttgct gtttgggtcc
     2941 cagcaacaca agttacatgt ctccctggga cactaataag ccccaaattg tcttccgctt
     3001 cacacaactc tacccagagg acagacatgg ggtggtgggg caggtggcgg ggagggggtg
     3061 gggcagtgca gtgaggggac gggtctccca acaagcagct gctgcaatgt ttccccaact
     3121 tcaagaagca gacactgggg ctgtgggcag agaggtaaga gtcagttggc ttgagagaac
     3181 tgctcttggc acagcctggc ccctgtcact tgagctggag tctgaagcac tgagaatttc
     3241 tgaccctcag gggaagcccc ctctcagcta tttgccctac tctgttcatt cgctgcctcc
     3301 tgtctatggc cccctcactc ctcccacctc tgcagactgg tagcatcact ccaggaacta
     3361 cattgagccc ttcacaaact gtaagcccta cacaactata gttaccattg caaagaaagc
     3421 taacctcgag atctgcgatc tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     3481 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atcgctgact
     3541 aatttttttt atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta
     3601 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag cttggtaccg cgggcccggg
     3661 atccaccggt cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca
     3721 tcctggtcga gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg
     3781 agggcgatgc cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc
     3841 ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct
     3901 accccgacca catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc
     3961 aggagcgcac catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt
     4021 tcgagggcga caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg
     4081 gcaacatcct ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg
     4141 ccgacaagca gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg
     4201 gcagcgtgca gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc
     4261 tgctgcccga caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga
     4321 agcgcgatca catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg
     4381 acgagctgta caagtaaagc ggccaattcc cccccccccc cccccctctc cctccccccc
     4441 ccctaacgtt actggccgaa gccgcttgga ataaggccgg tgtgcgtttg tctatatgtt
     4501 attttccacc atattgccgt cttttggcaa tgtgagggcc cggaaacctg gccctgtctt
     4561 cttgacgagc attcctaggg gtctttcccc tctcgccaaa ggaatgcaag gtctgttgaa
     4621 tgtcgtgaag gaagcagttc ctctggaagc ttcttgaaga caaacaacgt ctgtagcgac
     4681 cctttgcagg cagcggaacc ccccacctgg cgacaggtgc ctctgcggcc aaaagccacg
     4741 tgtataagat acacctgcaa aggcggcaca accccagtgc cacgttgtga gttggatagt
     4801 tgtggaaaga gtcaaatggc tctcctcaag cgtattcaac aaggggctga aggatgccca
     4861 gaaggtaccc cattgtatgg gatctgatct ggggcctcgg tgcacatgct ttacatgtgt
     4921 ttagtcgagg ttaaaaaaac gtctaggccc cccgaaccac ggggacgtgg ttttcctttg
     4981 aaaaacacga tgataatatg gggtcgaggc tgctgctgct gctgctgctg ggcctgaggc
     5041 tacagctctc cctgggcatc atcccagttg aggaggagaa cccggacttc tggaaccgcg
     5101 aggcagccga ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga
     5161 acctcatcat cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc
     5221 taaaagggca gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc
     5281 catatgtggc tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca
     5341 cagccacggc ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag
     5401 ccgcccgctt taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc
     5461 gggccaagaa agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct
     5521 cgccagccgg cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc
     5581 ctgcctcggc ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg
     5641 acattgacgt gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc
     5701 ctgagtaccc agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc
     5761 aggaatggct ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc
     5821 aggcttccct ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga
     5881 aatacgagat ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg
     5941 ccctgcgcct gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca
     6001 tcgaccatgg tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg
     6061 acgacgccat tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca
     6121 ctgccgacca ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct
     6181 tcgggctggc ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa
     6241 acggtccagg ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg
     6301 ggagccccga gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg
     6361 aggacgtggc ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc
     6421 agaccttcat agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg
     6481 acctggcgcc ccccgccggc accaccgacg ccgcgcaccc ggggcggtcc gtggtccccg
     6541 cgttgcttcc tctgctggcc gggaccctgc tgctgctgga gacggccact gctccctgag
     6601 tgtcccgtcc ctggggctcc tgcttcccca tcccggagtt ctcctgctcc ccacctcctg
     6661 tcgtcctgcc tggcctccag cccgagtcgt catccccgga gtccctatac agaggtcctg
     6721 ccatggaacc ttcccctccc cgtgcgctct ggggactgag cccatgacac caaacctgcc
     6781 ccttggctgc tctcggactc cctaccccaa ccccagggac tgcaggttgt gccctgtggc
     6841 tgcctgcacc ccaggaaagg agggggctca ggccatccag ccaccaccta cagcccagtg
     6901 gggtcgagga tatcgcggcc ggccgcactc ctcaggtgca ggctgcctat cagaaggtgg
     6961 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     7021 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     7081 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     7141 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     7201 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     7261 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     7321 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     7381 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     7441 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     7501 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     7561 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctc
//
