LOCUS       sequence-203026-        6560 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 6560)
  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 6560)
  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-EB3-GFP
FEATURES             Location/Qualifiers
     source          1..6560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     regulatory      1826..1835
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     regulatory      2690..2699
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(2696..2698,2699..2701,2702..3415)
                     /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(2696..2698,2699..2701,2702..3412)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2741..2762)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3002..3021)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3349..3370
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    3564..3685
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3601..3620)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3655..3674
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3692..4147)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3829..3850)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4041..4060
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4174..4278
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4280..4637
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4280..4471
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4303..4323)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4488..4623
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4672..5466
                     /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             4672..5466
                     /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(4726..4745)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5336..5355
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5654..5673)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5698..5745
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(6074..6560,1..102)
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
       61 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
      121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
      181 cctgattctg tggataaccg tattaccgcc atgcattagt tattactagc gctaccggac
      241 tcagatctcg agctcaagct tcgaattctg cagcccgggg gatccttcag gggtgtcgct
      301 tggttatttc caaaaatcaa attaatttta ttaaactatt agaacgagca tgttttgtct
      361 atatgctaca gaagataaaa aataatagga gttaacagtt ataaaacaac acactttgtt
      421 tctattgatt gttgaccaca ctggggtctc attaagttag attaaagaca cactaactgg
      481 gtcaaaagca gcagattgat ttcatagcac cagggtaaac tttctaacac ttttacggca
      541 atcatataca ttaaaattaa atacagacca cgactgaaca aggaggatga tctccaatat
      601 taaacaaaga gacttgtgcc tatttctctg agggtaaaca tgacctctca agttagcaag
      661 ttgtttttaa cactacaaaa atagttaaga ctgcaatccc agaataaagt attggtttta
      721 accaatcaat atagtacagt aaacatccat ttgttttgtt gaaacgttaa acaaatctga
      781 ccaaagctat tagcttatat aaaacaggtt tgccttctat gtagctgaaa acaccacagg
      841 cccgattttg ctactgtgta aaacatttca gcaagatttt tttattgcat ttttttttac
      901 tgaatcgttc aaacatttta tcattttagt ttgttcattc attgcaactg gaaaaacaac
      961 acatcacaca accgcacata tttcagcaat aagtacaata aaacactcaa ataaaaaaaa
     1021 cattttaaat ctctttgtat ttttgaccgc tgtttcgcgt aatttcacgg taaaactctg
     1081 gaaatctcca ctacattcct ctcagcggct cctctcaatg acagctgaag aagtgacgcg
     1141 gctgcctgct gtgttttgat tggtcgaatt cactggaggc ttccagaaca gtgtagagtc
     1201 tgaacgggtg cgcgctctgc tgtatttaaa gggcgaaaga gagaccgcag agaaactcaa
     1261 ccgaagagaa gcgacttgac aaagaagaaa agagcagcct gacaggactt tttccccgac
     1321 gaggtgttta ttcgctctat ttaagaatct actgtaaggt aagtctcaat atattgtact
     1381 ctattggcta atcagaatta tatagagatt atatgtactt aatgtcaaaa aatcaacttt
     1441 gtatatgtaa tctttttaca tgtggactgc ctatgttcat cttattttag gtctactaga
     1501 aaattatatt tcccgttttc acaataagga tttttaaaaa aagcaatgaa cagacgggca
     1561 tttactttat gttgctgaca ttattttata tgagcataat aaccataaat actagcaaat
     1621 gtcctaaatg aatttgtgtt aatgttgtct acaaaagaaa attagcgttt tacttgtaca
     1681 actaataata acttggttat taagagaatt tcacttgttg actagaaaaa tcctttcata
     1741 atgaaacaat tgcaccataa attgtataaa tataaaatta attctaattg tttttttttc
     1801 ctgcagtcga attctgcagt cgactgccac catggccgtc aatgtgtact ccacatctgt
     1861 gaccagtgaa aatctgagtc gccatgatat gcttgcatgg gtcaacgact ccctgcacct
     1921 caactatacc aagatagaac agctttgttc aggggcagcc tactgccagt tcatggacat
     1981 gctcttcccc ggctgtgtgc acttgaggaa agtgaagttc caggccaaac tagagcatga
     2041 atacatccac aacttcaagg tgctgcaagc agctttcaag aagatgggtg ttgacaaaat
     2101 cattcctgta gagaaattag tgaaaggaaa attccaagat aattttgagt ttattcagtg
     2161 gtttaagaaa ttctttgacg caaactatga tggaaaggat tacaaccctc tgctggcgcg
     2221 gcagggccag gacgtagcgc cacctcctaa cccaggtgat cagatcttca acaaatccaa
     2281 gaaactcatt ggcacagcag ttccacagag gacgtccccc acaggcccaa aaaacatgca
     2341 gacctctggc cggctgagca atgtggcccc cccctgcatt ctccggaaga atcctccatc
     2401 agcccgaaat ggcggccatg agactgatgc ccaaattctt gaactcaacc aacagctggt
     2461 ggacttgaag ctgacagtgg atgggctgga gaaggaacgt gacttctact tcagcaaact
     2521 tcgtgacatc gagctcatct gccaggagca tgaaagtgaa aacagccctg ttatctcagg
     2581 catcattggc atcctctatg ccacagagga aggattcgca ccccctgagg acgatgagat
     2641 tgaagagcat caacaagaag accaggacga gtacctggat ccaccggtcg ccaccatggt
     2701 gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga
     2761 cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa
     2821 gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt
     2881 gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca
     2941 cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa
     3001 ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa
     3061 ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct
     3121 ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga agaacggcat
     3181 caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca
     3241 ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct
     3301 gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct
     3361 ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg
     3421 cctcgagcct ctagcgaccc gggcggccgc gactctagat cataatcagc cataccacat
     3481 ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata
     3541 aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa
     3601 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3661 tgtccaaact catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa
     3721 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     3781 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     3841 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     3901 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     3961 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     4021 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     4081 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     4141 ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc
     4201 taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa
     4261 tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4441 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     4501 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     4561 aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     4621 gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa
     4681 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     4741 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     4801 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag
     4861 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     4921 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     4981 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5041 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5101 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5161 gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat
     5221 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5281 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5341 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     5401 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     5461 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     5521 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     5581 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta
     5641 gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg
     5701 caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc
     5761 ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc
     5821 gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag
     5881 ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg
     5941 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     6001 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     6061 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
     6121 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
     6181 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
     6241 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
     6301 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
     6361 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
     6421 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
     6481 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
     6541 agcgcacgag ggagcttcca
//
