LOCUS       sequence-41835-e        4579 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 4579)
  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 4579)
  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_mFGF_eGFP
FEATURES             Location/Qualifiers
     source          1..4579
                     /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"
     regulatory      510..519
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(516..518,519..521,522..1235)
                     /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(516..518,519..521,522..1232)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(561..582)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(822..841)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1169..1190
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1358..1479
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1395..1414)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1449..1468
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(1486..1941)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1623..1644)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     1835..1854
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        1968..2072
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2074..2431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2074..2265
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2097..2117)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2282..2417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2466..3260
                     /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             2466..3260
                     /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(2520..2539)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3130..3149
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3448..3467)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    3492..3539
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3868..4456
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4357..4376
                     /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 ccaccggtcg ccaccatggt gagcaagggc gaggagctgt
      541 tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc cacaagttca
      601 gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg aagttcatct
      661 gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg acctacggcg
      721 tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc aagtccgcca
      781 tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc aactacaaga
      841 cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag ctgaagggca
      901 tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac tacaacagcc
      961 acaacgtcta tatcatggcc gacaagcaga agaacggcat caaggtgaac ttcaagatcc
     1021 gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag aacaccccca
     1081 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag tccgccctga
     1141 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
     1201 ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct agatcataat
     1261 cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac acctccccct
     1321 gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg cagcttataa
     1381 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     1441 ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt gtaagcgtta
     1501 atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
     1561 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
     1621 ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa
     1681 aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg
     1741 ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt
     1801 gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg
     1861 ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta
     1921 atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     1981 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2041 aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc agctgtggaa
     2101 tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     2161 catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc cagcaggcag
     2221 aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc taactccgcc
     2281 catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct gactaatttt
     2341 ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga agtagtgagg
     2401 aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga tgaggatcgt
     2461 ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc
     2521 tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc
     2581 tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg
     2641 aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag
     2701 ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg
     2761 ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg
     2821 caatgcggcg gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac
     2881 atcgcatcga gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg
     2941 acgaagagca tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgagcatgc
     3001 ccgacggcga ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg
     3061 aaaatggccg cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc
     3121 aggacatagc gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc
     3181 gcttcctcgt gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc
     3241 ttcttgacga gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc
     3301 caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg
     3361 aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt
     3421 cttcgcccac cctaggggga ggctaactga aacacggaag gagacaatac cggaaggaac
     3481 ccgcgctatg acggcaataa aaagacagaa taaaacgcac ggtgttgggt cgtttgttca
     3541 taaacgcggg gttcggtccc agggctggca ctctgtcgat accccaccga gaccccattg
     3601 gggccaatac gcccgcgttt cttccttttc cccaccccac cccccaagtt cgggtgaagg
     3661 cccagggctc gcagccaacg tcggggcggc aggccctgcc atagcctcag gttactcata
     3721 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3781 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3841 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3901 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3961 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     4021 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     4081 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     4141 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     4201 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4261 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4321 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4381 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4441 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4501 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4561 cgccatgcat tagttatta
//
