LOCUS       sequence-203252-        9303 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 9303)
  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 9303)
  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-Par3-GFP
FEATURES             Location/Qualifiers
     source          1..9303
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1531..1549
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     1531..1548
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     regulatory      5012..5021
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5018..5020,5021..5023,5024..5734)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(5018..5020,5021..5023,5024..5734)
                     /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(5063..5084)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5324..5343)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     5671..5692
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    5785..5919
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5809..5828)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5863..5882
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(6032..6052)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(6032..6050)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(6071..6087)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6071..6087)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6084..6106)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6095..6111
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(join(6119..6125,6126..6143,6144..6149))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6164..6185
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6302..6319)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6473..7061)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6553..6572)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7232..8023,8024..8092))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     7855..7874
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8093..8197)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(8223..8678)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8360..8381)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8572..8591
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 tcaggggtgt cgcttggtta tttccaaaaa tcaaattaat tttattaaac tattagaacg
       61 agcatgtttt gtctatatgc tacagaagat aaaaaataat aggagttaac agttataaaa
      121 caacacactt tgtttctatt gattgttgac cacactgggg tctcattaag ttagattaaa
      181 gacacactaa ctgggtcaaa agcagcagat tgatttcata gcaccagggt aaactttcta
      241 acacttttac ggcaatcata tacattaaaa ttaaatacag accacgactg aacaaggagg
      301 atgatctcca atattaaaca aagagacttg tgcctatttc tctgagggta aacatgacct
      361 ctcaagttag caagttgttt ttaacactac aaaaatagtt aagactgcaa tcccagaata
      421 aagtattggt tttaaccaat caatatagta cagtaaacat ccatttgttt tgttgaaacg
      481 ttaaacaaat ctgaccaaag ctattagctt atataaaaca ggtttgcctt ctatgtagct
      541 gaaaacacca caggcccgat tttgctactg tgtaaaacat ttcagcaaga tttttttatt
      601 gcattttttt ttactgaatc gttcaaacat tttatcattt tagtttgttc attcattgca
      661 actggaaaaa caacacatca cacaaccgca catatttcag caataagtac aataaaacac
      721 tcaaataaaa aaaacatttt aaatctcttt gtatttttga ccgctgtttc gcgtaatttc
      781 acggtaaaac tctggaaatc tccactacat tcctctcagc ggctcctctc aatgacagct
      841 gaagaagtga cgcggctgcc tgctgtgttt tgattggtcg aattcactgg aggcttccag
      901 aacagtgtag agtctgaacg ggtgcgcgct ctgctgtatt taaagggcga aagagagacc
      961 gcagagaaac tcaaccgaag agaagcgact tgacaaagaa gaaaagagca gcctgacagg
     1021 actttttccc cgacgaggtg tttattcgct ctatttaaga atctactgta aggtaagtct
     1081 caatatattg tactctattg gctaatcaga attatataga gattatatgt acttaatgtc
     1141 aaaaaatcaa ctttgtatat gtaatctttt tacatgtgga ctgcctatgt tcatcttatt
     1201 ttaggtctac tagaaaatta tatttcccgt tttcacaata aggattttta aaaaaagcaa
     1261 tgaacagacg ggcatttact ttatgttgct gacattattt tatatgagca taataaccat
     1321 aaatactagc aaatgtccta aatgaatttg tgttaatgtt gtctacaaaa gaaaattagc
     1381 gttttacttg tacaactaat aataacttgg ttattaagag aatttcactt gttgactaga
     1441 aaaatccttt cataatgaaa caattgcacc ataaattgta taaatataaa attaattcta
     1501 attgtttttt tttcctgcag tcgaagcttg atttaggtga cactatagaa tacaagctac
     1561 ttgttctttt tgcaggatcc catcgattcg aattcgtgtt ggaactatga aagtgacggt
     1621 gtgttttggg agaactcggg tcgtggttcc gtgcggagac ggaaacatta aagtgcagtc
     1681 gctcgtccag caggccgcga tgcggtaccg gagagcgatc gccaagggtg aggagtactg
     1741 ggttcaggtt taccggctgg aacacggtga tggaggaatc ttggacctgg atgatgtgct
     1801 gtgtgatgtg gccgatgaca aagacagatt agtggcggtg tttgatgaac aggagcctca
     1861 tgttggagga gacggcacca gtgccagctc tacaggcaca cagtctcctg aactctactg
     1921 tggagaaccg tccacctcca ccccattgtc tgccttccaa ccctacctgc cccacagcga
     1981 gatagaagtc accacatcca cactgcgcac caatatgcct ttgcatgttc gccgcagtag
     2041 tgatccggca ttgttaaacc tgacggcgat gtctttttct gagccgggct cgcaacccga
     2101 agagccttcc aggaaaaacc ccacacgctg gtccaccact gctggcttcc tgaaaccacg
     2161 cttcgccact ggcaccaaca gcctggagag gaagggtcga ggtgtggaca catatcgtag
     2221 tcttcctcgt gatgcagggc agtggtccaa tcagaaagag tttcagcgtg aaaaggcccg
     2281 ttcttcactc agtgccaatc atcccatggt ggaccgctgg ctggagaggc aggagcagga
     2341 tgaggaggag aatggaagga tagaaccggt gggacgtgcg gacacatgct tggagcacat
     2401 gggagtgaga tcattggatg atattgtgaa attggtggaa gtgtccaatg acgggggtcc
     2461 actcggcatc catgttgtcc ccttcagtgg tcgagaccgc aggacgttgg gtctgctggt
     2521 gaagcgattg gagcgtggtg gaaaggctga tgtccagggt ttgtttcagg agaatgactg
     2581 catcattcgg atcaacaacg gagatctgcg caacgttcgc tttgagcaag ctcagaacat
     2641 gtttcgtcaa gcaatgcgtt ctcccgtcat cctgttccat gtggtgccaa cctccatgag
     2701 gtcccagtat gagcagatat cccataatga gcacaacccc agagcgaaca tggacctctc
     2761 gggacgtttc agcccagact ccctcaccaa tgatctggac agcgctgctc accgattggc
     2821 tcaacacaga ccacagccac ccaataacca tctggatact ggctctcccg tccatcatct
     2881 tgttggctca tcgggcaaac ctccgacagg ccatacttca tcaccccaaa gaggcctaag
     2941 ccccgccccc acaacaggct ttactaagaa ggttgggcgg agactcggaa tccagcttaa
     3001 gaaaggacct gaaggtctcg ggtttagcat cacgtcccga gacgttcctc ttggagggtc
     3061 tgcacctatt tatgtcaaga acattctgcc aagaggagct gccatacaag acggcagact
     3121 gaaagctgga gatagattgc tagaggtaaa tggggtggat ctgaatgggc ggggccagga
     3181 ggaggtggtc tctctgctaa gggcgacgcc catgggtggg actgtcactc tggtggtgct
     3241 gagacaggaa gagaccttta ttcccagaga gatgaacgca gaaccagcaa tacagaacac
     3301 aagagaatgg aaggtagaag aggaggaatt ggtcttgact ccggatggta cgcgggagtt
     3361 tctgacatta gaagtgcctc tgaatgattc tggctcagca ggattaggag tcagtgtcaa
     3421 gggtaacagg tcaaaggaaa gccacgctga cctgggcatc ttcgtcaagt ctattatcaa
     3481 tggaggagca gcttgcaagg acggaaggct gcgggtcaac gatcagctta ttgcagtcaa
     3541 tggggagtcg ttattaggga agacaaacca ggatgccatg gaaacgctgc gcaagtccat
     3601 gtcaacggaa ggcaacaagc gagggatgat tcagctgatc gtggcgagac gcatcaacaa
     3661 gcgcctcgag ggtgagagcc gcagcagtcc tcgagggtta gagcgaacac tgagcccgtc
     3721 tcctgatgac catgagcggc gaatttcaca ctcactgtat gggatcgagg gccttgatga
     3781 caacctgagg ccacaagcca gcacaaataa ccgcaaaatg aatcattatc agctctcgcc
     3841 gacggtgaac atgcctcagg acgacacggt cattattgaa gatgacagac cacacgttct
     3901 ccccatccag ctgtctgacc aatcctcgtc cagttctcat gatgacatgg gtttcgccgt
     3961 ggagacaccc ccgccgccac cctgggaacc agagcttccc gacagctctt ccagtgctaa
     4021 tgctgagggg cagtttcaga gggaaggctt tggcaggcag agcatgtctg agaaacgcac
     4081 taagcagtat ggagatgcag gccaactcga catcatcaag acccgcaagt ccaagagcat
     4141 ggacctagtg gctgatgaga ttaacctcac acaatgcaca gagaaccaca caggctcttc
     4201 cacgcgtgat gtgggcccgt ctctgggact gaagaagtcc agctctctgg aaagtttaca
     4261 gaccgcagtc gcagaggtta cgctcaatgg tgacatgccg tttcaccggc cgcgaccacg
     4321 gattataaga ggacgaggct gcaatgagag tttcagagct gccattgaca aatcctacga
     4381 caggcctgca gccaatgagg atgaagagga gtgcatggac acactggagg aggacactga
     4441 aggaagctct cgatccggcc gggattcagt ttccaccgtg gccgatctca ctccactccc
     4501 tgtcaccgag caacagctga tcaatggaaa ccagcctgaa aacgacaaga agaaagaaaa
     4561 aggggggaag gacaaaaaga agccggagaa agagaagggt aaaacaaaga aaggaatgct
     4621 caagggcctc ggagagatgt tcaggtttgg aaagtaccgt aaggatgagc gattagacgg
     4681 tgccaagtgg aaagcagagg aaacccatgc atctgaagag gagactcgca gaatgaagca
     4741 agagcaggag aggatccagg caaaaacgcg agagattcgt cagcgtcagg ccagagagcg
     4801 agactatgcc gagatccagg acttcagtcg atccaccctg acctcacttc ctcccgagga
     4861 gcctccgtac gcaggcatcg gctctctgga gcacggcggg taccatcgca ttcacactcc
     4921 tccagattca ccatacacac agaagcagaa cggacgcaat ggacacccct ccacttcaga
     4981 caggtacgtc gacggtaccg cgggcccggt cgccaccatg gtgagcaagg gcgaggagct
     5041 gttcaccggg gtggtgccca tcctggtcga gctggacggc gacgtaaacg gccacaagtt
     5101 cagcgtgtcc ggcgagggcg agggcgatgc cacctacggc aagctgaccc tgaagttcat
     5161 ctgcaccacc ggcaagctgc ccgtgccctg gcccaccctc gtgaccaccc tgacctacgg
     5221 cgtgcagtgc ttcagccgct accccgacca catgaagcag cacgacttct tcaagtccgc
     5281 catgcccgaa ggctacgtcc aggagcgcac catcttcttc aaggacgacg gcaactacaa
     5341 gacccgcgcc gaggtgaagt tcgagggcga caccctggtg aaccgcatcg agctgaaggg
     5401 catcgacttc aaggaggacg gcaacatcct ggggcacaag ctggagtaca actacaacag
     5461 ccacaacgtc tatatcatgg ccgacaagca gaagaacggc atcaaggtga acttcaagat
     5521 ccgccacaac atcgaggacg gcagcgtgca gctcgccgac cactaccagc agaacacccc
     5581 catcggcgac ggccccgtgc tgctgcccga caaccactac ctgagcaccc agtccgccct
     5641 gagcaaagac cccaacgaga agcgcgatca catggtcctg ctggagttcg tgaccgccgc
     5701 cgggatcact ctcggcatgg acgagctgta caagtccgga ctcagatctc gagcctctag
     5761 aactatagtg agtcgtatta cgtagatcca gacatgataa gatacattga tgagtttgga
     5821 caaaccacaa ctagaatgca gtgaaaaaaa tgctttattt gtgaaatttg tgatgctatt
     5881 gctttatttg taaccattat aagctgcaat aaacaagtta acaacaacaa ttgcattcat
     5941 tttatgtttc aggttcaggg ggaggtgtgg gaggtttttt aattcgcggc cgcggcgcca
     6001 atgcattggg cccggtaccc agcttttgtt ccctttagtg agggttaatt gcgcgcttgg
     6061 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     6121 acatacgagc cgggagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     6181 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc
     6241 attaatgaat cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt
     6301 cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact
     6361 caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag
     6421 caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata
     6481 ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc
     6541 cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg
     6601 ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc
     6661 tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg
     6721 gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc
     6781 ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga
     6841 ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg
     6901 gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa
     6961 aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg
     7021 tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt
     7081 ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat
     7141 tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct
     7201 aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta
     7261 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     7321 ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac
     7381 gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     7441 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     7501 taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg
     7561 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     7621 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     7681 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     7741 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     7801 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     7861 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     7921 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     7981 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     8041 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atactcttcc
     8101 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     8161 aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga aaagtgccac
     8221 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     8281 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     8341 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     8401 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     8461 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     8521 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     8581 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     8641 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
     8701 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagt cgaccgccaa
     8761 ttcaatatgg cgtatatgga ctcatgccaa ttcaatatgg tggatctgga cctgtgccaa
     8821 ttcaatatgg cgtatatgga ctcgtgccaa ttcaatatgg tggatctgga ccccagccaa
     8881 ttcaatatgg cggacttggc accatgccaa ttcaatatgg cggacctggc actgtgccaa
     8941 ctggggaggg gtctacttgg cacggtgcca agtttgagga ggggtcttgg ccctgtgcca
     9001 agtccgccat attgaattgg catggtgcca ataatggcgg ccatattggc tatatgccag
     9061 gatcaatata taggcaatat ccaatatggc cctatgccaa tatggctatt ggccaggttc
     9121 aatactatgt attggcccta tgccatatag tattccatat atgggttttc ctattgacgt
     9181 agatagcccc tcccaatggg cggtcccata taccatatat ggggcttcct aataccgccc
     9241 atagccactc ccccattgac gtcaatggtc tctatatatg gtctttccta ttgacgtcat
     9301 atg
//
