LOCUS       sequence-457676-        7337 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes the AURKA biosensor where LC3B is flanked by the 
            Aquamarine/TdLanYFP donor/acceptor FRET pair and under the control 
            of the CMV promoter..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7337)
  AUTHORS   Bousmah Y, Valenta H, Bertolin G, Singh U, Nicolas V, Pasquier H, 
            Tramier M, Merola F, Erard M
  TITLE     tdLanYFP, a Yellow, Bright, Photostable, and pH-Insensitive 
            Fluorescent Protein for Live-Cell Imaging and Forster Resonance 
            Energy Transfer-Based Sensing Strategies.
  JOURNAL   ACS Sens. 2021 Nov 26;6(11):3940-3947. doi: 
            10.1021/acssensors.1c00874. Epub 2021 Oct 22.
  PUBMED    34676768
REFERENCE   2  (bases 1 to 7337)
  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: 
            "10.1021/acssensors.1c00874"; journalName: "ACS Sens"; date: 
            "2021-11-26- 26"; volume: "6"; issue: "11"; pages: "3940-3947"
COMMENT     Sequence Label: TdLanYFP-AURKA-Aquamarine
FEATURES             Location/Qualifiers
     source          1..7337
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      53..188
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             237..1031
                     /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             237..1031
                     /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(291..310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     complement(1219..1238)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    1263..1310
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1639..2227
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2128..2147
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        2402..2705
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2706..2909
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2860..2880
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             2987..3646
                     /codon_start=1
                     /product="tetrameric yellow fluorescent protein from 
                     Branchiostoma lanceolatum, also known as blFP-Y3"
                     /label=LanYFP
                     /translation="MASLPATHELHIFGSFNGVDFDMVGRGTGNPNDGYEELNLKSTKG
                     DLQFSPWILVPQIGYGFHQYLPFPDGMSPFQAAMKDGSGYQVHRTMQFEDGASLTSNYR
                     YTYEGSHIKGEFQVKGTGFPADGPVMTNSLTAADWCVTKMLYPNDKTIISTFDWTYTTG
                     NGKRYQSTARTTYTFAKPMAANILKNQPMFVFRKTELKHSKTELNFKEWQKAFTDVM"
     CDS             3698..4357
                     /codon_start=1
                     /product="tetrameric yellow fluorescent protein from 
                     Branchiostoma lanceolatum, also known as blFP-Y3"
                     /label=LanYFP
                     /translation="MASLPATHELHIFGSFNGVDFDMVGRGTGNPNDGYEELNLKSTKG
                     DLQFSPWILVPQIGYGFHQYLPFPDGMSPFQAAMKDGSGYQVHRTMQFEDGASLTSNYR
                     YTYEGSHIKGEFQVKGTGFPADGPVMTNSLTAADWCVTKMLYPNDKTIISTFDWTYTTG
                     NGKRYQSTARTTYTFAKPMAANILKNQPMFVFRKTELKHSKTELNFKEWQKAFTDVM"
     CDS             5294..5305
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      5618..5627
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(5624..5626,5627..5629,5630..6343)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5669..5690)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(5930..5949)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6277..6298
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    6466..6587
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6503..6522)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6557..6576
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(6594..7049)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6731..6752)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6943..6962
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        7076..7180
                     /gene="bla"
                     /label=AmpR promoter
     promoter        join(7182..7337,1..202)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        join(7182..7337,1..36)
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(7205..7225)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
       61 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
      121 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
      181 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
      241 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
      301 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
      361 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
      421 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
      481 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
      541 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
      601 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
      661 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
      721 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
      781 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
      841 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
      901 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
      961 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     1021 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     1081 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     1141 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     1201 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     1261 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     1321 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     1381 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     1441 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     1501 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     1561 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     1621 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     1681 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     1741 tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc tagtgtagcc
     1801 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     1861 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     1921 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     1981 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     2041 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     2101 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     2161 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     2221 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     2281 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     2341 ttagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     2401 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     2461 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     2521 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     2581 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     2641 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     2701 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     2761 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     2821 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     2881 tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc cgctagcgct
     2941 accggactca gatctcgaga tggtctccaa aggagaggag gataacatgg cctctctccc
     3001 agcgacacat gagttacaca tctttggctc cttcaacggt gtggactttg acatggtggg
     3061 tcgtggcacc ggcaatccaa atgatggtta tgaggagtta aacctgaagt ccaccaaggg
     3121 tgacctccag ttctccccct ggattctggt ccctcaaatc gggtatggat tccatcagta
     3181 cctgcccttc cccgacggga tgtcgccttt ccaggccgcc atgaaagatg gctccggata
     3241 ccaagtccat cgcacaatgc agtttgaaga cggtgcctcc ctgacttcca actaccgcta
     3301 cacctacgag ggaagccaca tcaaaggaga gtttcaggtg aaggggactg gtttccctgc
     3361 tgacggtcct gtgatgacca actcgctgac cgctgcggac tggtgcgtga ccaagatgct
     3421 gtaccccaac gacaaaacca tcatcagcac ctttgactgg acttacacca ctggaaatgg
     3481 caagcgctac cagagcactg cgcggaccac ctacaccttt gccaagccaa tggcggccaa
     3541 catcctgaag aaccagccga tgttcgtgtt ccgtaagacg gaactcaagc actccaagac
     3601 cgaactcaac ttcaaggagt ggcaaaaggc atttaccgat gtgatgtcga ctggcactgg
     3661 ttctacgggc tcgggctcct caggagagga ggataacatg gcctctctcc cagcgacaca
     3721 tgagttacac atctttggct ccttcaacgg tgtggacttt gacatggtgg gtcgtggcac
     3781 cggcaatcca aatgatggtt atgaggagtt aaacctgaag tccaccaagg gtgacctcca
     3841 gttctccccc tggattctgg tccctcaaat cgggtatggc ttccatcagt acctgccctt
     3901 ccccgacggg atgtcgcctt tccaggccgc catgaaagat ggctccggat accaagtcca
     3961 tcgcacaatg cagtttgaag acggtgcctc cctgacttcc aactaccgct acacctacga
     4021 gggaagccac atcaaaggag agtttcaggt gaaggggact ggtttccctg ctgacggtcc
     4081 tgtgatgacc aactcgctga ccgctgcgga ctggtgcgtg accaagatgc tgtaccccaa
     4141 cgacaaaacc atcatcagca cctttgactg gacttacacc actggaaatg gcaagcgcta
     4201 ccagagcact gcgcggacca cctacacctt tgccaagcca atggcggcca acatcctgaa
     4261 gaaccagccg atgttcgtgt tccgtaagac ggaactcaag cactccaaga ccgaactcaa
     4321 cttcaaggag tggcaaaagg catttaccga tgtgatgggc atggacgagc tgtacaaggg
     4381 aagcttcgaa ttcatggacc gatctaaaga aaactgcatt tcaggacctg ttaaggctac
     4441 agctccagtt ggaggtccaa aacgtgttct cgtgactcag caatttcctt gtcagaatcc
     4501 attacctgta aatagtggcc aggctcagcg ggtcttgtgt ccttcaaatt cttcccagcg
     4561 cattcctttg caagcacaaa aacttgtctc cagtcacaag ccggttcaga atcagaagca
     4621 gaagcaattg caggcaacca gtgtacctca tcctgtctcc aggccactga ataacaccca
     4681 aaagagcaag cagcccctgc catcggcacc tgaaaataat cctgaggagg aactggcatc
     4741 aaaacagaaa aatgaagaat caaaaaagag gcagtgggct ttggaagact ttgaaattgg
     4801 tcgccctctg ggtaaaggaa agtttggtaa tgtttatttg gcaagagaaa agcaaagcaa
     4861 gtttattctg gctcttaaag tgttatttaa agctcagctg gagaaagccg gagtggagca
     4921 tcagctcaga agagaagtag aaatacagtc ccaccttcgg catcctaata ttcttagact
     4981 gtatggttat ttccatgatg ctaccagagt ctacctaatt ctggaatatg caccacttgg
     5041 aacagtttat agagaacttc agaaactttc aaagtttgat gagcagagaa ctgctactta
     5101 tataacagaa ttggcaaatg cactatcata ttgccattcg aagagagtta ttcatagaga
     5161 cattaagcca gagaacttac ttcttggatc agctggagag cttaaaattg cagattttgg
     5221 gtggtcagta catgctccat cttccaggag gaccactctc tgtggcaccc tggactacct
     5281 gccccctgaa atgattgaag gtcggatgca tgatgagaag gtggatctct ggagccttgg
     5341 agttctttgc tatgaatttt tagttgggaa gcctcctttt gaggcaaaca cataccaaga
     5401 gacctacaaa agaatatcac gggttgagtt cacattccct gactttgtaa cagagggagc
     5461 cagggacctc atttcaagac tgttgaagca taatcccagc cagaggccaa tgctcagaga
     5521 agtacttgaa cacccctgga tcacagcaaa ttcatcaaaa ccatcaaatt gccaaaacaa
     5581 agaatcagct agcaaacagt ctggggatcc accggtcgcc accatggtga gcaagggcga
     5641 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
     5701 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
     5761 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgag
     5821 ctggggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
     5881 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
     5941 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     6001 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     6061 catcagcggc aacgtctata tcaccgccga caagcagaag aacggcatca aggccaactt
     6121 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     6181 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     6241 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     6301 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcgactctag
     6361 atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac
     6421 ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca
     6481 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6541 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt
     6601 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6661 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6721 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6781 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6841 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6901 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     6961 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7021 cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg
     7081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     7141 accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag
     7201 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     7261 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     7321 gcaggcagaa gtatgca
//
