LOCUS       sequence-203025-        7532 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 7532)
  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 7532)
  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-aPKC-CAAX-GFP
FEATURES             Location/Qualifiers
     source          1..7532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     3..22
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             join(1811..1813,1814..1816,1817..2527)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(1811..1813,1814..1816,1817..2527)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1856..1877)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2117..2136)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2464..2485
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4536..4657
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4573..4592)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4627..4646
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4664..5119)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4801..4822)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5013..5032
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5146..5250
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5252..5609
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5252..5443
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5275..5295)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5460..5595
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             5644..6438
                     /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             5644..6438
                     /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(5698..5717)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6308..6327
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(6626..6645)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    6670..6717
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      join(7046..7532,1..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 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt
     1861 cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga
     1921 tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc
     1981 ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga
     2041 ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg
     2101 caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg
     2161 cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat
     2221 cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa
     2281 gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt
     2341 gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc
     2401 cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga
     2461 tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct
     2521 gtacaagtcc ggactcagat ctcgagctca agcttcgaat tcgatgccca gcaggacgga
     2581 ccccaagatg gaccggagcg gcggccgcgt ccgtctgaag gcgcactacg gcggggacat
     2641 cctgattacc agcgtggatg ccatgacaac attcaaggac ctctgtgagg aagtgcgaga
     2701 catgtgtggc ctgcaccagc agcacccact caccctcaag tgggtggaca gtgaaggtga
     2761 cccttgtact gtgtcctcac agatggagct ggaggccttc cgcctggcct gtcagggcag
     2821 ggacgaagtg ctcatcatcc acgttttccc aagcatccca gaacaaccgg gcatgccttg
     2881 tcctggagaa gacaagtcca tctaccgccg tggagccaga agatggagga agctataccg
     2941 agccaacggc cacctcttcc aagccaagcg ctttaacagg agagcgtact gtggccagtg
     3001 cagcgaaagg atatggggcc tcgcgaggca ggggtacagg tgcatcaact gcaagctgct
     3061 tgtccataaa cgctgccacg tcctcgtccc gctgacctgc aggaggcata tggattctgt
     3121 catgccttcc caagagcctc cagtagatga caagaacgat ggtgtagacc ttccttcaga
     3181 agaaactgat ggaattgctt atatttcttc atctcggaaa catgacaata tcaaagatga
     3241 ttctgaggac cttaagcctg tcatcgatgg ggtggatggg atcaaaatct ctcaggggct
     3301 ggggctgcaa gactttgacc tcatcagagt catcgggcgt ggaagctatg ccaaggtcct
     3361 cctggtgcgg ttgaagaaaa acgaccagat ttacgccatg aaggtggtga agaaggagct
     3421 cgtccacgac gatgaggata tcgactgggt gcagacagag aagcacgtgt tcgagcaggc
     3481 atccagcaac cccttcctgg ttggcttaca ctcctgcttc cagacaacga gccggttgtt
     3541 cctggtcatc gagtatgtca acggggggga cctcatgttc cacatgcaga ggcagaggaa
     3601 gcttccagag gaacacgcca ggttctatgc tgctgagatc tgtatcgctc tcaacttcct
     3661 acatgagaga gggatcatct accgggacct aaaactggac aacgtcctcc tcgatgccga
     3721 tggacacatt aagctgacgg actacggcat gtgcaaggaa ggcctaggcc ccggcgacac
     3781 aacaagcact ttttgtggaa ccccgaacta tatcgccccc gaaatcctgc gaggagaaga
     3841 gtacgggttc agcgtggact ggtgggcgct gggtgtcctt atgtttgaga tgatggctgg
     3901 gcgctccccc tttgacatca tcacagacaa ccctgacatg aatactgaag actacctttt
     3961 ccaagttatc ctggaaaagc caattcggat tccccgtttc ctgtctgtca aggcctcaca
     4021 cgtcttgaaa ggatttttaa ataaggatcc caaagagagg cttggctgcc ggccgcagac
     4081 tgggttttcc gacatcaagt cccatgcctt cttccgaagc atagactggg acctgcttga
     4141 aaagaagcag accctgcctc ccttccagcc ccagatcaca gatgactatg gcctggacaa
     4201 cttcgacacg cagttcacca gcgagcccgt acagctgacc ccagatgatg aggacgtcat
     4261 aaagaggatc gaccagtccg agttcgaagg cttcgagtac atcaacccgc ttctgctgtc
     4321 tgctgaggag tccgtgctcg agaagctgaa ccctcctgat gagagtggcc ccggctgcat
     4381 gagctgcaag tgtgtgctct cctaatcgac ccgggcggcc gcgactctag atcataatca
     4441 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     4501 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     4561 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     4621 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     4681 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     4741 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     4801 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     4861 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     4921 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     4981 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     5041 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     5101 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     5161 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     5221 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     5281 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     5341 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5401 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     5461 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     5521 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     5581 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     5641 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5701 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     5761 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     5821 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     5881 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     5941 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     6001 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     6061 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     6121 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     6181 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     6241 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     6301 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     6361 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     6421 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     6481 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     6541 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     6601 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     6661 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     6721 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     6781 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     6841 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     6901 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     6961 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     7021 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     7081 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     7141 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     7201 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     7261 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     7321 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     7381 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     7441 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     7501 tcggaacagg agagcgcacg agggagcttc ca
//
