LOCUS       sequence-446951-        7531 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 7531)
  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 7531)
  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 mR0-37 +SV40 base prom-GFP-IRES-AP
FEATURES             Location/Qualifiers
     source          1..7531
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(788..807)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    853..908
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(907..926)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(1269..1285)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1269..1285)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1282..1304)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1293..1309
                     /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(1317..1323,1324..1341,1342..1347))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1362..1383
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        1442..1637
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      1488..1623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    1643..1777
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1680..1699)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1734..1753
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1845..1862)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2016..2604)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2096..2115)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2775..3566,3567..3635))
                     /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     3398..3417
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3636..3740)
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4755..4951
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4802..4937
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     regulatory      4986..4995
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(4992..4994,4995..4997,4998..5711)
                     /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(4992..4994,4995..4997,4998..5708)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5037..5058)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5298..5317)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5645..5666
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    5738..6311
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    5761..6313
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(5905..5922)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     6132..6151
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     CDS             join(6325..7531,1..293)
                     /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"
ORIGIN
        1 cctacacggt cctcctatac ggaaacggtc caggctatgt gctcaaggac ggcgcccggc
       61 cggatgttac cgagagcgag agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc
      121 tggacgaaga gacccacgca ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc
      181 acctggttca cggcgtgcag gagcagacct tcatagcgca cgtcatggcc ttcgccgcct
      241 gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc
      301 acccggggcg gtccgtggtc cccgcgttgc ttcctctgct ggccgggacc ctgctgctgc
      361 tggagacggc cactgctccc tgagtgtccc gtccctgggg ctcctgcttc cccatcccgg
      421 agttctcctg ctccccacct cctgtcgtcc tgcctggcct ccagcccgag tcgtcatccc
      481 cggagtccct atacagaggt cctgccatgg aaccttcccc tccccgtgcg ctctggggac
      541 tgagcccatg acaccaaacc tgccccttgg ctgctctcgg actccctacc ccaaccccag
      601 ggactgcagg ttgtgccctg tggctgcctg caccccagga aaggaggggg ctcaggccat
      661 ccagccacca cctacagccc agtggggtcg aggatatcgc ggccggccgc actcctcagg
      721 tgcaggctgc ctatcagaag gtggtggctg gtgtggccaa tgccctggct cacaaatacc
      781 actgagatct ttttccctct gccaaaaatt atggggacat catgaagccc cttgagcatc
      841 tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg aattttttgt
      901 gtctctcact cggaaggaca tatgggaggg caaatcattt aaaacatcag aatgagtatt
      961 tggtttagag tttggcaaca tatgcccata tgctggctgc catgaacaaa ggttggctat
     1021 aaagaggtca tcagtatatg aaacagcccc ctgctgtcca ttccttattc catagaaaag
     1081 ccttgacttg aggttagatt ttttttatat tttgttttgt gttatttttt tctttaacat
     1141 ccctaaaatt ttccttacat gttttactag ccagattttt cctcctctcc tgactactcc
     1201 cagtcatagc tgtccctctt ctcttatgga gatccctcga cctgcagccc aagcttggcg
     1261 taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat tccacacaac
     1321 atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag ctaactcaca
     1381 ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg ccagcggatc
     1441 cgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     1501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     1561 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     1621 gcctaggctt ttgcaaaaag ctaacttgtt tattgcagct tataatggtt acaaataaag
     1681 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1741 gtccaaactc atcaatgtat cttatcatgt ctggatccgc tgcattaatg aatcggccaa
     1801 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     1861 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     1921 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     1981 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2041 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2101 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2161 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2221 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2281 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2341 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2401 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2461 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2521 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2581 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2641 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2701 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2761 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     2821 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     2881 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     2941 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3001 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3061 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3121 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3181 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3241 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3301 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3361 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3421 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3481 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3541 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3601 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3661 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3721 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgggtc gacactaggt
     3781 ctaactggct gttcatatct gtttagacct ttgccacttt ggaagtgtgc ccctatgtct
     3841 gaaaaggcca ttgaccttcc caccttttgt ctggtagatt ttgttttaga ccagggacag
     3901 ttggagactt caggagatac caagtgcaac tgcgagtccc ctggttggct ggagtgctgg
     3961 cagagcctgt taattagcac aagaacaccc caatgctcca ttagccaggc caagcattcg
     4021 gaacttcccc tcctggggcc tcttcctcct aatgagagca ggagaagggc tcttacaaaa
     4081 cattgcctta gaataatatt ctgctttgtc ccccaccagt aaccagggtt gacaggatca
     4141 ctctgtccat ttctcctcta attagcacaa ttccagcatg gcctgagcca gcaaatggag
     4201 aaaagcgatc aaaagcgatc tgacaaccgt accgaatcac aaatacaaag agagccattc
     4261 cctgtaatta ggtgagacaa tccctcagtc ctttctcggc tggagtttgc tgctaagact
     4321 cttgctgttt gggtcccagc agcacagtta catgtctccc tgggacacta ataagcccca
     4381 gattgtcttc cgcttcatac aactctaacc aggggacaga tgcagggagg agggatgggg
     4441 aacacggtct ctaaacaaag caactcgtga gaggtttccc aaagcccaaa aggcacaggc
     4501 tgtggagcag agcagaactg cactggcaca gcctaggctc catccctgga gccagagtct
     4561 gaagtactgg agctctgacc ttcctggaag cccgctctgt ttccttacct ccagccctca
     4621 ctccttgcat ggcctgcagc atggtgactg cggtaccacg gtgactgagg tacgacaccc
     4681 agacctccat acccagacct gcagtgaggt tcttaccacc atgtcaaaga gaatttacct
     4741 cgagatctgc gatctgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     4801 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatcgct gactaatttt
     4861 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     4921 aggctttttt ggaggcctag gcttttgcaa aaagcttggt accgcgggcc cgggatccac
     4981 cggtcgccac catggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
     5041 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
     5101 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
     5161 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
     5221 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     5281 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     5341 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     5401 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     5461 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     5521 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     5581 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     5641 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     5701 tgtacaagta aagcggccaa ttcccccccc cccccccccc ctctccctcc ccccccccta
     5761 acgttactgg ccgaagccgc ttggaataag gccggtgtgc gtttgtctat atgttatttt
     5821 ccaccatatt gccgtctttt ggcaatgtga gggcccggaa acctggccct gtcttcttga
     5881 cgagcattcc taggggtctt tcccctctcg ccaaaggaat gcaaggtctg ttgaatgtcg
     5941 tgaaggaagc agttcctctg gaagcttctt gaagacaaac aacgtctgta gcgacccttt
     6001 gcaggcagcg gaacccccca cctggcgaca ggtgcctctg cggccaaaag ccacgtgtat
     6061 aagatacacc tgcaaaggcg gcacaacccc agtgccacgt tgtgagttgg atagttgtgg
     6121 aaagagtcaa atggctctcc tcaagcgtat tcaacaaggg gctgaaggat gcccagaagg
     6181 taccccattg tatgggatct gatctggggc ctcggtgcac atgctttaca tgtgtttagt
     6241 cgaggttaaa aaaacgtcta ggccccccga accacgggga cgtggttttc ctttgaaaaa
     6301 cacgatgata atatggggtc gaggctgctg ctgctgctgc tgctgggcct gaggctacag
     6361 ctctccctgg gcatcatccc agttgaggag gagaacccgg acttctggaa ccgcgaggca
     6421 gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc caagaacctc
     6481 atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag gatcctaaaa
     6541 gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg cttcccatat
     6601 gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg agccacagcc
     6661 acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag tgcagccgcc
     6721 cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat gaatcgggcc
     6781 aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca cgcctcgcca
     6841 gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga cgtgcctgcc
     6901 tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa catggacatt
     6961 gacgtgatcc taggtggagg ccgaaagtac atgtttcgca tgggaacccc agaccctgag
     7021 tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct ggtgcaggaa
     7081 tggctggcga agcgccaggg tgcccggtat gtgtggaacc gcactgagct catgcaggct
     7141 tccctggacc cgtctgtgac ccatctcatg ggtctctttg agcctggaga catgaaatac
     7201 gagatccacc gagactccac actggacccc tccctgatgg agatgacaga ggctgccctg
     7261 cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac
     7321 catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat gttcgacgac
     7381 gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct cgtcactgcc
     7441 gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc catcttcggg
     7501 ctggcccctg gcaaggcccg ggacaggaag g
//
