LOCUS       sequence-41721-e        5818 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5818)
  AUTHORS   Bestman JE, Lee-Osbourne J, Cline HT
  TITLE     In vivo time-lapse imaging of cell proliferation and differentiation
            in the optic tectum of Xenopus laevis tadpoles.
  JOURNAL   J Comp Neurol. 2012 Feb 1;520(2):401-33. doi: 10.1002/cne.22795.
  PUBMED    22113462
REFERENCE   2  (bases 1 to 5818)
  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: "10.1002/cne"; 
            journalName: "J Comp Neurol"; date: "2012-02-1- 1"; volume: "520"; 
            issue: "2"; pages: "401-33"
COMMENT     Sequence Label: pSox2-bd::FP
FEATURES             Location/Qualifiers
     source          1..5818
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     49..65
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     49..65
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             503..943
                     /codon_start=1
                     /gene="Saccharomyces cerevisiae GAL4 (truncated)"
                     /product="DNA binding domain of the GAL4 transcriptional 
                     activator"
                     /label=GAL4 DNA binding domain
                     /translation="MKLLSSIEQACDICRLKKLKCSKEKPKCAKCLKNNWECRYSPKTK
                     RSPLTRAHLTEVESRLERLEQLFLLIFPREDLDMILKMDSLQDIKALLTGLFVQDNVNK
                     DAVTDRLASVETDMPLTLRQHRISATSSSEESSNKGQRQLTVS"
     regulatory      1749..1758
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1755..1757,1758..1760,1761..2474)
                     /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(1755..1757,1758..1760,1761..2471)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1800..1821)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2061..2080)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2408..2429
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2597..2718
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2634..2653)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2688..2707
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2725..3180)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2862..2883)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3074..3093
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3207..3311
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3313..3670
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3313..3504
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3336..3356)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3521..3656
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3705..4499
                     /codon_start=1
                     /product="aminoglycoside phosphotransferase"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     CDS             3705..4499
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(3759..3778)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4369..4388
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4687..4706)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4731..4778
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5107..5695
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5596..5615
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ctagcgctac cggactcaga tctcgagctc caccgcggtg gcggccgctc tagaactagt
       61 ggatctgaaa actctttgtt tggatgctaa tgggatactt aaaggatctg aaaactcttt
      121 gtttggatgc taatgggata cttaaaggat ctgaaaactc tttgtttgga tgctaatggg
      181 atacttaaag gatctgaaaa ctctttgttt ggatgctaat gggatactta aaggatctga
      241 aaactctttg tttggatgct aatgggatac ttaaaggatc tgaaaactct ttgtttggat
      301 gctaatggga tacttaaagg atcccccggg cgcggggcag gcgagtagga gggggcgccg
      361 gctatatata ccactgctcc ggagggctgg gcgcgcgggg actatcccgc caccgttgcg
      421 tccctatttg ctctcgctac ttaggtctgt gcgcagcact caccgaactc acggcccgca
      481 gctcgaactc acgccaagat ccatgaagct actgtcttct atcgaacaag catgcgatat
      541 ttgccgactt aaaaagctca agtgctccaa agaaaaaccg aagtgcgcca agtgtctgaa
      601 gaacaactgg gagtgtcgct actctcccaa aaccaaaagg tctccgctga ctagggcaca
      661 tctgacagaa gtggaatcaa ggctagaaag actggaacag ctatttctac tgatttttcc
      721 tcgagaagac cttgacatga ttttgaaaat ggattcttta caggatataa aagcattgtt
      781 aacaggatta tttgtacaag ataatgtgaa taaagatgcc gtcacagata gattggcttc
      841 agtggagact gatatgcctc taacattgag acagcataga ataagtgcga catcatcatc
      901 ggaagagagt agtaacaaag gtcaaagaca gttgactgta tcgtcgaggt cgaccccggg
      961 aattcagatc tctcgagccg cccccccgac cgatgtcagc ctgggggacg agctccactt
     1021 agacggcgag gacgtggcga tggcgcatgc cgacgcgcta gacgatttcg atctggacat
     1081 gttgggggac ggggattccc cgggtccggg atttaccccc cacgactccg ccccctacgg
     1141 cgctctggat atgtagagat ctactagtct cgagtctaga atcaagctta ggcctccaag
     1201 gcggagtact gtctccgggc tggcggagta ctgtcctccg gcaaggtcgg agtactgtcc
     1261 tccgacacta gaggtcggag tactgtcctc cgacgcaagg cggagtactg tcctccgggc
     1321 tgcggagtac tgtcctccgg caaggtcgga gtactgtcct ccgacactag aggtcggagt
     1381 actgtcctcc gacgcaaggt cggagtactg tcctccgaca ctagaggtcg gagtactgtc
     1441 ctccgacgca aggtcggagt actgtcctcc gacactagag gtcggagtac tgtcctccga
     1501 cgcaaggcgg agtactgtcc tccgggctgg cggagtactg tcctccggca agggtcgaga
     1561 tcccccgggc gcggggcagg cgagtaggag ggggcgccgg ctatatatac cactgctccg
     1621 gagggctggg cgcgcgggga ctatcccgcc accgttgcgt ccctatttgc tctcgctact
     1681 taggtctgtg cgcagcactc accgaactca cggcccgcag ctcgaactca cgccaagatc
     1741 caccggtcgc caccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1801 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1861 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1921 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1981 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     2041 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     2101 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     2161 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     2221 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     2281 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     2341 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     2401 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     2461 agctgtacaa gtaaagcggc cgcgactcta gatcataatc agccatacca catttgtaga
     2521 ggttttactt gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa
     2581 tgcaattgtt gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag
     2641 catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa
     2701 actcatcaat gtatcttaag gcgtaaattg taagcgttaa tattttgtta aaattcgcgt
     2761 taaatttttg ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt
     2821 ataaatcaaa agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc
     2881 cactattaaa gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg
     2941 gcccactacg tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac
     3001 taaatcggaa ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg
     3061 tggcgagaaa ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag
     3121 cggtcacgct gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt
     3181 caggtggcac ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac
     3241 attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa
     3301 aaaggaagag tcctgaggcg gaaagaacca gctgtggaat gtgtgtcagt tagggtgtgg
     3361 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     3421 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     3481 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     3541 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     3601 ggccgcctcg gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     3661 cttttgcaaa gatcgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     3721 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     3781 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     3841 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaagac gaggcagcgc
     3901 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     3961 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     4021 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     4081 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     4141 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     4201 cgccagccga actgttcgcc aggctcaagg cgagcatgcc cgacggcgag gatctcgtcg
     4261 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     4321 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     4381 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     4441 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     4501 cgggactctg gggttcgaaa tgaccgacca agcgacgccc aacctgccat cacgagattt
     4561 cgattccacc gccgccttct atgaaaggtt gggcttcgga atcgttttcc gggacgccgg
     4621 ctggatgatc ctccagcgcg gggatctcat gctggagttc ttcgcccacc ctagggggag
     4681 gctaactgaa acacggaagg agacaatacc ggaaggaacc cgcgctatga cggcaataaa
     4741 aagacagaat aaaacgcacg gtgttgggtc gtttgttcat aaacgcgggg ttcggtccca
     4801 gggctggcac tctgtcgata ccccaccgag accccattgg ggccaatacg cccgcgtttc
     4861 ttccttttcc ccaccccacc ccccaagttc gggtgaaggc ccagggctcg cagccaacgt
     4921 cggggcggca ggccctgcca tagcctcagg ttactcatat atactttaga ttgatttaaa
     4981 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     5041 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     5101 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     5161 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     5221 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     5281 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     5341 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     5401 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     5461 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     5521 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     5581 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     5641 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     5701 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     5761 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gccatgcatt agttatta
//
