LOCUS       sequence-209756-        6327 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  measure LATS activity by Luciferase Assay.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6327)
  AUTHORS   Azad T, Janse van Rensburg HJ, Lightbody ED, Neveu B, Champagne A, 
            Ghaffari A, Kay VR, Hao Y, Shen H, Yeung B, Croy BA, Guan KL, 
            Pouliot F, Zhang J, Nicol CJB, Yang X
  TITLE     A LATS biosensor screen identifies VEGFR as a regulator of the Hippo
            pathway in angiogenesis.
  JOURNAL   Nat Commun. 2018 Mar 13;9(1):1061. doi: 10.1038/s41467-018-03278-w.
  PUBMED    29535383
REFERENCE   2  (bases 1 to 6327)
  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"; journalName: "Nat 
            Commun."; date: "2018-03-13"; volume: "9(1):1061. doi"; pages: " 
            10.1038/s41467-018-03278-w"
COMMENT     Sequence Label: pCDNA3.1neo-NLucYAP15
FEATURES             Location/Qualifiers
     source          1..6327
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      704..713
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(776..794)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2065..2082)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2071..2295
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      complement(2427..3015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2507..2526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     polyA_signal    3344..3391
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     primer_bind     3416..3435
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     CDS             complement(3623..4417)
                     /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             complement(3623..4417)
                     /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(3734..3753)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     4344..4363
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(4452..4809)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4466..4601
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     enhancer        4618..4809
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     4766..4786
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     CDS             complement(join(4869..5660,5661..5729))
                     /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     5492..5511
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5730..5834)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      5861..6316
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5948..5967)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     6158..6179
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacttaagc ttggtaccga gctcggatcc gccgccacca tggaagacgc
      721 caaaaacata aagaaaggcc cggcgccatt ctatccgctg gaagatggaa ccgctggaga
      781 gcaactgcat aaggctatga agagatacgc cctggttcct ggaacaattg cttttacaga
      841 tgcacatatc gaggtggaca tcacttacgc tgagtacttc gaaatgtccg ttcggttggc
      901 agaagctatg aaacgatatg ggctgaatac aaatcacaga atcgtcgtat gcagtgaaaa
      961 ctctcttcaa ttctttatgc cggtgttggg cgcgttattt atcggagttg cagttgcgcc
     1021 cgcgaacgac atttataatg aacgtgaatt gctcaacagt atgggcattt cgcagcctac
     1081 cgtggtgttc gtttccaaaa aggggttgca aaaaattttg aacgtgcaaa aaaagctccc
     1141 aatcatccaa aaaattatta tcatggattc taaaacggat taccagggat ttcagtcgat
     1201 gtacacgttc gtcacatctc atctacctcc cggttttaat gaatacgatt ttgtgccaga
     1261 gtccttcgat agggacaaga caattgcact gatcatgaac tcctctggat ctactggtct
     1321 gcctaaaggt gtcgctctgc ctcatagaac tgcctgcgtg agattctcgc atgccagaga
     1381 tcctattttt ggcaatcaaa tcattccgga tactgcgatt ttaagtgttg ttccattcca
     1441 tcacggtttt ggaatgttta ctacactcgg atatttgata tgtggatttc gagtcgtctt
     1501 aatgtataga tttgaagaag agctgtttct gaggagcctt caggattaca agattcaaag
     1561 tgcgctgctg gtgccaaccc tattctcctt cttcgccaaa agcactctga ttgacaaata
     1621 cgatttatct aatttacacg aaattgcttc tggtggcgct cccctctcta aggaagtcgg
     1681 ggaagcggtt gccaagaggt tccatctgcc aggtatcagg caaggatatg ggctcactga
     1741 gactacatca gctattctga ttacacccga gggggatgat aaaccgggcg cggtcggtaa
     1801 agttgttcca ttttttgaag cgaaggttgt ggatctggat accgggaaaa cgctgggcgt
     1861 taatcaaaga ggcgaactgt gtgtgagagg tcctatgatt atgtccggtt atgtaaacaa
     1921 tccggaagcg accaacgcct tgattgacaa ggatggagga ggagctggag gcccacagca
     1981 tgttcgagct cattcctctc cagcttctct gcagttgtaa gcggccgctc gagtctagag
     2041 ggcccgttta aacccgctga tcagcctcga ctgtgccttc tagttgccag ccatctgttg
     2101 tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct
     2161 aataaaatga ggaaattgca tcgcattgtc tgagtaggtg tcattctatt ctggggggtg
     2221 gggtggggca ggacagcaag ggggaggatt gggaagacaa tagcaggcat gctggggatg
     2281 cggtgggctc tatggcttct gaggcggaaa gaaccagtgg cggtaatacg gttatccaca
     2341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     2401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     2461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     2521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     2581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     2641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     3001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     3061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     3121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta acctgaggct
     3181 atggcagggc ctgccgcccc gacgttggct gcgagccctg ggccttcacc cgaacttggg
     3241 gggtggggtg gggaaaagga agaaacgcgg gcgtattggc cccaatgggg tctcggtggg
     3301 gtatcgacag agtgccagcc ctgggaccga accccgcgtt tatgaacaaa cgacccaaca
     3361 ccgtgcgttt tattctgtct ttttattgcc gtcatagcgc gggttccttc cggtattgtc
     3421 tccttccgtg tttcagttag cctcccccta gggtgggcga agaactccag catgagatcc
     3481 ccgcgctgga ggatcatcca gccggcgtcc cggaaaacga ttccgaagcc caacctttca
     3541 tagaaggcgg cggtggaatc gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc
     3601 atttcgaacc ccagagtccc gctcagaaga actcgtcaag aaggcgatag aaggcgatgc
     3661 gctgcgaatc gggagcggcg ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc
     3721 caagctcttc agcaatatca cgggtagcca acgctatgtc ctgatagcgg tccgccacac
     3781 ccagccggcc acagtcgatg aatccagaaa agcggccatt ttccaccatg atattcggca
     3841 agcaggcatc gccatgggtc acgacgagat cctcgccgtc gggcatgctc gccttgagcc
     3901 tggcgaacag ttcggctggc gcgagcccct gatgctcttc gtccagatca tcctgatcga
     3961 caagaccggc ttccatccga gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga
     4021 atgggcaggt agccggatca agcgtatgca gccgccgcat tgcatcagcc atgatggata
     4081 ctttctcggc aggagcaagg tgagatgaca ggagatcctg ccccggcact tcgcccaata
     4141 gcagccagtc ccttcccgct tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg
     4201 tcgtggccag ccacgatagc cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca
     4261 ggtcggtctt gacaaaaaga accgggcgcc cctgcgctga cagccggaac acggcggcat
     4321 cagagcagcc gattgtctgt tgtgcccagt catagccgaa tagcctctcc acccaagcgg
     4381 ccggagaacc tgcgtgcaat ccatcttgtt caatcatgcg aaacgatcct catcctgtct
     4441 cttgatcgat ctttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat
     4501 agctcagagg ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg
     4561 ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg
     4621 actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
     4681 ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat acttctgcct
     4741 gctggggagc ctggggactt tccacaccct aactgacaca cattccacag ctggttcttt
     4801 ccgcctcagg actcttcctt tttcaataaa tcaatctaaa gtatatatga gtaaacttgg
     4861 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     4921 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     4981 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5041 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5101 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5161 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5221 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5281 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5341 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5401 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5461 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5521 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5581 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5641 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     5701 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     5761 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     5821 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgcgccctg tagcggcgca
     5881 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     5941 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     6001 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     6061 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     6121 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     6181 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     6241 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata
     6301 ttaacgctta caatttacgc gcgcgtt
//
