LOCUS       sequence-446987-        7065 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 7065)
  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 7065)
  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 mR0-37a +SV40 base prom-GFP-IRES-AP
FEATURES             Location/Qualifiers
     source          1..7065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(25..46)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     374..393
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     633..654
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      696..705
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(740..936)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      754..889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     promoter        1487..1591
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1592..1660,1661..2452)
                     /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     complement(1810..1829)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2623..3211
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3112..3131
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3365..3382
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3450..3584
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3474..3493)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3528..3547
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(3590..3785)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3604..3739
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    3844..3865
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3880..3885,3886..3903,3904..3910)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3918..3934
                     /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)."
     primer_bind     3923..3945
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3942..3958
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3942..3958
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4301..4320
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4319..4374
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4420..4439
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(4934..6433)
                     /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"
     misc_feature    6445..6997
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    6447..7020
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(6607..6626)
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     6836..6853
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     CDS             complement(join(7045..7065,1..693,694..696,697..699))
                     /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             complement(join(7048..7065,1..693,694..696,697..699))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
ORIGIN
        1 gccgagagtg atcccggcgg cggtcacgaa ctccagcagg accatgtgat cgcgcttctc
       61 gttggggtct ttgctcaggg cggactgggt gctcaggtag tggttgtcgg gcagcagcac
      121 ggggccgtcg ccgatggggg tgttctgctg gtagtggtcg gcgagctgca cgctgccgtc
      181 ctcgatgttg tggcggatct tgaagttcac cttgatgccg ttcttctgct tgtcggccat
      241 gatatagacg ttgtggctgt tgtagttgta ctccagcttg tgccccagga tgttgccgtc
      301 ctccttgaag tcgatgccct tcagctcgat gcggttcacc agggtgtcgc cctcgaactt
      361 cacctcggcg cgggtcttgt agttgccgtc gtccttgaag aagatggtgc gctcctggac
      421 gtagccttcg ggcatggcgg acttgaagaa gtcgtgctgc ttcatgtggt cggggtagcg
      481 gctgaagcac tgcacgccgt aggtcagggt ggtcacgagg gtgggccagg gcacgggcag
      541 cttgccggtg gtgcagatga acttcagggt cagcttgccg taggtggcat cgccctcgcc
      601 ctcgccggac acgctgaact tgtggccgtt tacgtcgccg tccagctcga ccaggatggg
      661 caccaccccg gtgaacagct cctcgccctt gctcaccatg gtggcgaccg gtggatcccg
      721 ggcccgcggt accaagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
      781 tctggaatag ctcagaggcc gaggcggcct cggcctctgc ataaataaaa aaaattagtc
      841 agcgatgggg cggagaatgg gcggaactgg gcggagttag gggcgggatg ggcggagtta
      901 ggggcgggac tatggttgct gactaattga gatgcagatc gcagatctcg aggaccgtgt
      961 tccccatccc tcctccctgc atctgtcccc tggttagagt tgtatgaagc ggaagacaat
     1021 ctggggctta ttagtgtccc agggagacat gtaactgtgc tgctgggacc caaacagcaa
     1081 gagtcttagc agcaaactcc agccgagaaa ggactgaggg attgtctcac ctaattacag
     1141 ggaatggctc tctttgtatt tgtgattcgg tacggttgtc agatcgcttt tgatcgcttt
     1201 tctccatttg ctggctcagg ccatgctgga attgtgctaa ttagaggaga aatggacaga
     1261 gtgatcctgt caaccctggt tactggtggg ggacaaagca gaatattatt ctaaggcaat
     1321 gttttgtaag agcccttctc ctgctctcat taggaggaag aggccccagg aggggaagtt
     1381 ccgaatgctt ggcctggcta atggagcatt ggggtgttct tgtgctaatt aacaggctct
     1441 gccagcactc caggtcgacc caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     1501 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     1561 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     1621 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     1681 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     1741 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     1801 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     1861 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     1921 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     1981 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     2041 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     2101 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     2161 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     2221 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     2281 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     2341 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     2401 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     2461 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     2521 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     2581 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     2641 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     2701 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     2761 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     2821 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     2881 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     2941 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     3001 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     3061 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     3121 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     3181 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     3241 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     3301 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     3361 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     3421 cgcgcgttgg ccgattcatt aatgcagcgg atccagacat gataagatac attgatgagt
     3481 ttggacaaac cacaactaga atgcagtgaa aaaaatgctt tatttgtgaa atttgtgatg
     3541 ctattgcttt atttgtaacc attataagct gcaataaaca agttagcttt ttgcaaaagc
     3601 ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggcc gaggcggcct
     3661 cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg
     3721 gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga
     3781 gatgcggatc cgctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3841 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     3901 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     3961 tgattacgcc aagcttgggc tgcaggtcga gggatctcca taagagaaga gggacagcta
     4021 tgactgggag tagtcaggag aggaggaaaa atctggctag taaaacatgt aaggaaaatt
     4081 ttagggatgt taaagaaaaa aataacacaa aacaaaatat aaaaaaaatc taacctcaag
     4141 tcaaggcttt tctatggaat aaggaatgga cagcaggggg ctgtttcata tactgatgac
     4201 ctctttatag ccaacctttg ttcatggcag ccagcatatg ggcatatgtt gccaaactct
     4261 aaaccaaata ctcattctga tgttttaaat gatttgccct cccatatgtc cttccgagtg
     4321 agagacacaa aaaattccaa cacactattg caatgaaaat aaatttcctt tattagccag
     4381 aagtcagatg ctcaaggggc ttcatgatgt ccccataatt tttggcagag ggaaaaagat
     4441 ctcagtggta tttgtgagcc agggcattgg ccacaccagc caccaccttc tgataggcag
     4501 cctgcacctg aggagtgcgg ccggccgcga tatcctcgac cccactgggc tgtaggtggt
     4561 ggctggatgg cctgagcccc ctcctttcct ggggtgcagg cagccacagg gcacaacctg
     4621 cagtccctgg ggttggggta gggagtccga gagcagccaa ggggcaggtt tggtgtcatg
     4681 ggctcagtcc ccagagcgca cggggagggg aaggttccat ggcaggacct ctgtataggg
     4741 actccgggga tgacgactcg ggctggaggc caggcaggac gacaggaggt ggggagcagg
     4801 agaactccgg gatggggaag caggagcccc agggacggga cactcaggga gcagtggccg
     4861 tctccagcag cagcagggtc ccggccagca gaggaagcaa cgcggggacc acggaccgcc
     4921 ccgggtgcgc ggcgtcggtg gtgccggcgg ggggcgccag gtcgcaggcg gtgtagggct
     4981 ccaggcaggc ggcgaaggcc atgacgtgcg ctatgaaggt ctgctcctgc acgccgtgaa
     5041 ccaggtgcgc ctgcgggccg cgcgcgaaca ccgccacgtc ctcgcctgcg tgggtctctt
     5101 cgtccagggg cactgctgac tgctgccgat actcggggct cccgctctcg ctctcggtaa
     5161 catccggccg ggcgccgtcc ttgagcacat agcctggacc gtttccgtat aggaggaccg
     5221 tgtaggcctt cctgtcccgg gccttgccag gggccagccc gaagatggag ctccctcgca
     5281 gggggtagcc tccgaaggag aagacgtggg agtggtcggc agtgacgagg ctcagcgtgt
     5341 cctcctcgct ggtgagctgg cccgccctct caatggcgtc gtcgaacatg atcgtctcag
     5401 tcagtgcccg gtaagccctg ctttcatgat gaccatggtc gatgcgacca ccctccacga
     5461 agaggaagaa gccgcggggg ttcctgctca gcaggcgcag ggcagcctct gtcatctcca
     5521 tcagggaggg gtccagtgtg gagtctcggt ggatctcgta tttcatgtct ccaggctcaa
     5581 agagacccat gagatgggtc acagacgggt ccagggaagc ctgcatgagc tcagtgcggt
     5641 tccacacata ccgggcaccc tggcgcttcg ccagccattc ctgcaccaga ttcttcccgt
     5701 ccagcctggt cccaccttgg ctgtagtcat ctgggtactc agggtctggg gttcccatgc
     5761 gaaacatgta ctttcggcct ccacctagga tcacgtcaat gtccatgttg gagatgagct
     5821 gcgtagcgat gtcctggcac ccctcctggc gggccgaggc aggcacgtcg gcgtccgagt
     5881 accagttgcg gttcaccgtg tgggcgtagg tgccggctgg cgaggcgtgc tgcactcgtg
     5941 tggtggttac cactcccact gacttccctg ctttcttggc ccgattcatc acggagatga
     6001 cctcgttgcc gcgtgtcgtg ttgcactggt taaagcgggc ggctgcactc aagccaatgg
     6061 tctggaagtt gcccttgacc ccgcacaggt aggccgtggc tgtggctcca ctgtctggca
     6121 catgtttgtc tacattgtat gtcttggaca gagccacata tgggaagcgg tccatggcca
     6181 ggggtatctc aggccccagt ttgtccttct tctgcccttt taggatcctg gcagctgtca
     6241 ccgtagacac ccccatccca tcgcccagga agatgatgag gttcttggcg gctgtctgtg
     6301 caggctgcag cttcttggcg gcacccaggg cctcggctgc ctcgcggttc cagaagtccg
     6361 ggttctcctc ctcaactggg atgatgccca gggagagctg tagcctcagg cccagcagca
     6421 gcagcagcag cagcctcgac cccatattat catcgtgttt ttcaaaggaa aaccacgtcc
     6481 ccgtggttcg gggggcctag acgttttttt aacctcgact aaacacatgt aaagcatgtg
     6541 caccgaggcc ccagatcaga tcccatacaa tggggtacct tctgggcatc cttcagcccc
     6601 ttgttgaata cgcttgagga gagccatttg actctttcca caactatcca actcacaacg
     6661 tggcactggg gttgtgccgc ctttgcaggt gtatcttata cacgtggctt ttggccgcag
     6721 aggcacctgt cgccaggtgg ggggttccgc tgcctgcaaa gggtcgctac agacgttgtt
     6781 tgtcttcaag aagcttccag aggaactgct tccttcacga cattcaacag accttgcatt
     6841 cctttggcga gaggggaaag acccctagga atgctcgtca agaagacagg gccaggtttc
     6901 cgggccctca cattgccaaa agacggcaat atggtggaaa ataacatata gacaaacgca
     6961 caccggcctt attccaagcg gcttcggcca gtaacgttag ggggggggga gggagagggg
     7021 gggggggggg cggaattggc cgctttactt gtacagctcg tccat
//
