LOCUS       sequence-203022-        6511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  heat shock inducible transgenesis.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6511)
  AUTHORS   Strzyz PJ, Lee HO, Sidhaye J, Weber IP, Leung LC, Norden C
  TITLE     Interkinetic nuclear migration is centrosome independent and ensures
            apical cell division to maintain tissue integrity.
  JOURNAL   Dev Cell. 2015 Jan 26;32(2):203-19. doi: 
            10.1016/j.devcel.2014.12.001. Epub 2015 Jan 15.
  PUBMED    25600237
REFERENCE   2  (bases 1 to 6511)
  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: 
            "10.1016/j.devcel.2014.12.001"; journalName: "Dev Cell"; date: 
            "2015-01-26- 26"; volume: "32"; issue: "2"; pages: "203-19"
COMMENT     Sequence Label: HS-PCNA-GFP
FEATURES             Location/Qualifiers
     source          1..6511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      1715..1724
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1721..1723,1724..1726,1727..2437)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1766..1787)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2027..2046)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2374..2395
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3413..3534
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3450..3469)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3504..3523
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3541..3996)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3678..3699)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3890..3909
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4023..4127
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4129..4486
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4129..4320
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4152..4172)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4337..4472
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4521..5315
                     /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             4521..5315
                     /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(4575..4594)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5185..5204
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5503..5522)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5547..5594
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5923..6511
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6412..6431
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
       61 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat gcattagtta
      121 ttactagcgc taccggactc agatctcgag ctcaagcttc gaattctgca gcccggggga
      181 tccttcaggg gtgtcgcttg gttatttcca aaaatcaaat taattttatt aaactattag
      241 aacgagcatg ttttgtctat atgctacaga agataaaaaa taataggagt taacagttat
      301 aaaacaacac actttgtttc tattgattgt tgaccacact ggggtctcat taagttagat
      361 taaagacaca ctaactgggt caaaagcagc agattgattt catagcacca gggtaaactt
      421 tctaacactt ttacggcaat catatacatt aaaattaaat acagaccacg actgaacaag
      481 gaggatgatc tccaatatta aacaaagaga cttgtgccta tttctctgag ggtaaacatg
      541 acctctcaag ttagcaagtt gtttttaaca ctacaaaaat agttaagact gcaatcccag
      601 aataaagtat tggttttaac caatcaatat agtacagtaa acatccattt gttttgttga
      661 aacgttaaac aaatctgacc aaagctatta gcttatataa aacaggtttg ccttctatgt
      721 agctgaaaac accacaggcc cgattttgct actgtgtaaa acatttcagc aagatttttt
      781 tattgcattt ttttttactg aatcgttcaa acattttatc attttagttt gttcattcat
      841 tgcaactgga aaaacaacac atcacacaac cgcacatatt tcagcaataa gtacaataaa
      901 acactcaaat aaaaaaaaca ttttaaatct ctttgtattt ttgaccgctg tttcgcgtaa
      961 tttcacggta aaactctgga aatctccact acattcctct cagcggctcc tctcaatgac
     1021 agctgaagaa gtgacgcggc tgcctgctgt gttttgattg gtcgaattca ctggaggctt
     1081 ccagaacagt gtagagtctg aacgggtgcg cgctctgctg tatttaaagg gcgaaagaga
     1141 gaccgcagag aaactcaacc gaagagaagc gacttgacaa agaagaaaag agcagcctga
     1201 caggactttt tccccgacga ggtgtttatt cgctctattt aagaatctac tgtaaggtaa
     1261 gtctcaatat attgtactct attggctaat cagaattata tagagattat atgtacttaa
     1321 tgtcaaaaaa tcaactttgt atatgtaatc tttttacatg tggactgcct atgttcatct
     1381 tattttaggt ctactagaaa attatatttc ccgttttcac aataaggatt tttaaaaaaa
     1441 gcaatgaaca gacgggcatt tactttatgt tgctgacatt attttatatg agcataataa
     1501 ccataaatac tagcaaatgt cctaaatgaa tttgtgttaa tgttgtctac aaaagaaaat
     1561 tagcgtttta cttgtacaac taataataac ttggttatta agagaatttc acttgttgac
     1621 tagaaaaatc ctttcataat gaaacaattg caccataaat tgtataaata taaaattaat
     1681 tctaattgtt tttttttcct gcagtcgacc ggtcgccacc atggtgagca agggcgagga
     1741 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     1801 gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     1861 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     1921 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     1981 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     2041 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     2101 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     2161 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg ccaacttcaa
     2221 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     2281 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     2341 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     2401 cgccgggatc actctcggca tggacgagct gtacaagggc gaagggcaag ggcaagggca
     2461 agggccgggc cgcggctacg cgtatcgatc catgttcgag gcgcgcctgg tccagggctc
     2521 catcctcaag aaggtgttgg aggcactcaa ggacctcatc aacgaggcct gctgggatat
     2581 tagctccagc ggtgtaaacc tgcagagcat ggactcgtcc cacgtctctt tggtgcagct
     2641 caccctgcgg tctgagggct tcgacaccta ccgctgcgac cgcaacctgg ccatgggcgt
     2701 gaacctcacc agtatgtcca aaatactaaa atgcgccggc aatgaagata tcattacact
     2761 aagggccgaa gataacgcgg ataccttggc gctagtattt gaagcaccaa accaggagaa
     2821 agtttcagac tatgaaatga agttgatgga tttagatgtt gaacaacttg gaattccaga
     2881 acaggagtac agctgtgtag taaagatgcc ttctggtgaa tttgcacgta tatgccgaga
     2941 tctcagccat attggagatg ctgttgtaat ttcctgtgca aaagacggag tgaaattttc
     3001 tgcaagtgga gaacttggaa atggaaacat taaattgtca cagacaagta atgtcgataa
     3061 agaggaggaa gctgttacca tagagatgaa tgaaccagtt caactaactt ttgcactgag
     3121 gtacctgaac ttctttacaa aagccactcc actctcttca acggtgacac tcagtatgtc
     3181 tgcagatgta ccccttgttg tagagtataa aattgcggat atgggacact taaaatacta
     3241 cttggctccc aagatcgagg atgaagaagg atcttagtct agtcgacccg ggcggccgcg
     3301 actctagatc ataatcagcc ataccacatt tgtagaggtt ttacttgctt taaaaaacct
     3361 cccacacctc cccctgaacc tgaaacataa aatgaatgca attgttgttg ttaacttgtt
     3421 tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc
     3481 atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttaaggcgt
     3541 aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat
     3601 tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga
     3661 tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca
     3721 acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct
     3781 aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc
     3841 cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag
     3901 cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca
     3961 cacccgccgc gcttaatgcg ccgctacagg gcgcgtcagg tggcactttt cggggaaatg
     4021 tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga
     4081 gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtcct gaggcggaaa
     4141 gaaccagctg tggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccagcagg
     4201 cagaagtatg caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg
     4261 ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc
     4321 gcccctaact ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca
     4381 tggctgacta atttttttta tttatgcaga ggccgaggcc gcctcggcct ctgagctatt
     4441 ccagaagtag tgaggaggct tttttggagg cctaggcttt tgcaaagatc gatcaagaga
     4501 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4561 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4621 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4681 ccggtgccct gaatgaactg caagacgagg cagcgcggct atcgtggctg gccacgacgg
     4741 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4801 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4861 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4921 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4981 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5041 tcaaggcgag catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5101 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5161 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5221 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5281 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5341 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5401 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5461 tctcatgctg gagttcttcg cccaccctag ggggaggcta actgaaacac ggaaggagac
     5521 aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa cgcacggtgt
     5581 tgggtcgttt gttcataaac gcggggttcg gtcccagggc tggcactctg tcgatacccc
     5641 accgagaccc cattggggcc aatacgcccg cgtttcttcc ttttccccac cccacccccc
     5701 aagttcgggt gaaggcccag ggctcgcagc caacgtcggg gcggcaggcc ctgccatagc
     5761 ctcaggttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5821 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5881 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5941 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     6001 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     6061 aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     6121 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     6181 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     6241 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     6301 cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     6361 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     6421 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     6481 atgctcgtca ggggggcgga gcctatggaa a
//
