LOCUS       sequence-457339-        5919 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Encodes a donor-only, Aquamarine-tagged version of the AURKA 
            biosensor under the control of a CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5919)
  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 5919)
  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: AURKA-Aquamarine
FEATURES             Location/Qualifiers
     source          1..5919
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(599..618)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    643..690
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      1019..1607
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1508..1527
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        1782..2085
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2086..2289
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2240..2260
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3256..3267
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     regulatory      3580..3589
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(3586..3588,3589..3591,3592..4305)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLSWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISGNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3631..3652)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3892..3911)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4239..4260
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4428..4549
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4465..4484)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4519..4538
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(4556..5011)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4693..4714)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4905..4924
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5038..5142
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5144..5501
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5144..5335
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5167..5187)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5352..5487
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             join(5536..5919,1..411)
                     /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             join(5536..5919,1..411)
                     /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(5590..5609)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
ORIGIN
        1 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
       61 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
      121 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
      181 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
      241 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
      301 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
      361 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
      421 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
      481 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
      541 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
      601 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
      661 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
      721 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
      781 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
      841 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
      901 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
      961 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     1021 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     1081 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     1141 gcagagcgca gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca
     1201 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     1261 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     1321 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     1381 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     1441 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     1501 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     1561 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     1621 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     1681 tatcccctga ttctgtggat aaccgtatta ccgccatgca ttagttatta atagtaatca
     1741 attacggggt cattagttca tagcccatat atggagttcc gcgttacata acttacggta
     1801 aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat aatgacgtat
     1861 gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga gtatttacgg
     1921 taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc ccctattgac
     1981 gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt atgggacttt
     2041 cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat gcggttttgg
     2101 cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag tctccacccc
     2161 attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc aaaatgtcgt
     2221 aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga ggtctatata
     2281 agcagagctg gtttagtgaa ccgtcagatc cgctagcgct accggactca gatctcgagc
     2341 tcaagcttcg aattcatgga ccgatctaaa gaaaactgca tttcaggacc tgttaaggct
     2401 acagctccag ttggaggtcc aaaacgtgtt ctcgtgactc agcaatttcc ttgtcagaat
     2461 ccattacctg taaatagtgg ccaggctcag cgggtcttgt gtccttcaaa ttcttcccag
     2521 cgcattcctt tgcaagcaca aaaacttgtc tccagtcaca agccggttca gaatcagaag
     2581 cagaagcaat tgcaggcaac cagtgtacct catcctgtct ccaggccact gaataacacc
     2641 caaaagagca agcagcccct gccatcggca cctgaaaata atcctgagga ggaactggca
     2701 tcaaaacaga aaaatgaaga atcaaaaaag aggcagtggg ctttggaaga ctttgaaatt
     2761 ggtcgccctc tgggtaaagg aaagtttggt aatgtttatt tggcaagaga aaagcaaagc
     2821 aagtttattc tggctcttaa agtgttattt aaagctcagc tggagaaagc cggagtggag
     2881 catcagctca gaagagaagt agaaatacag tcccaccttc ggcatcctaa tattcttaga
     2941 ctgtatggtt atttccatga tgctaccaga gtctacctaa ttctggaata tgcaccactt
     3001 ggaacagttt atagagaact tcagaaactt tcaaagtttg atgagcagag aactgctact
     3061 tatataacag aattggcaaa tgcactatca tattgccatt cgaagagagt tattcataga
     3121 gacattaagc cagagaactt acttcttgga tcagctggag agcttaaaat tgcagatttt
     3181 gggtggtcag tacatgctcc atcttccagg aggaccactc tctgtggcac cctggactac
     3241 ctgccccctg aaatgattga aggtcggatg catgatgaga aggtggatct ctggagcctt
     3301 ggagttcttt gctatgaatt tttagttggg aagcctcctt ttgaggcaaa cacataccaa
     3361 gagacctaca aaagaatatc acgggttgag ttcacattcc ctgactttgt aacagaggga
     3421 gccagggacc tcatttcaag actgttgaag cataatccca gccagaggcc aatgctcaga
     3481 gaagtacttg aacacccctg gatcacagca aattcatcaa aaccatcaaa ttgccaaaac
     3541 aaagaatcag ctagcaaaca gtctggggat ccaccggtcg ccaccatggt gagcaagggc
     3601 gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc tggacggcga cgtaaacggc
     3661 cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca cctacggcaa gctgaccctg
     3721 aagttcatct gcaccaccgg caagctgccc gtgccctggc ccaccctcgt gaccaccctg
     3781 agctggggcg tgcagtgctt cagccgctac cccgaccaca tgaagcagca cgacttcttc
     3841 aagtccgcca tgcccgaagg ctacgtccag gagcgcacca tcttcttcaa ggacgacggc
     3901 aactacaaga cccgcgccga ggtgaagttc gagggcgaca ccctggtgaa ccgcatcgag
     3961 ctgaagggca tcgacttcaa ggaggacggc aacatcctgg ggcacaagct ggagtacaac
     4021 tacatcagcg gcaacgtcta tatcaccgcc gacaagcaga agaacggcat caaggccaac
     4081 ttcaagatcc gccacaacat cgaggacggc agcgtgcagc tcgccgacca ctaccagcag
     4141 aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagcacccag
     4201 tccgccctga gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg
     4261 accgccgccg ggatcactct cggcatggac gagctgtaca agtaaagcgg ccgcgactct
     4321 agatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     4381 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     4441 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     4501 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttaa ggcgtaaatt
     4561 gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag ctcatttttt
     4621 aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg
     4681 ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga ctccaacgtc
     4741 aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc accctaatca
     4801 agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg gagcccccga
     4861 tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa
     4921 ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc
     4981 gccgcgctta atgcgccgct acagggcgcg tcaggtggca cttttcgggg aaatgtgcgc
     5041 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     5101 taaccctgat aaatgcttca ataatattga aaaaggaaga gtcctgaggc ggaaagaacc
     5161 agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc aggctcccca gcaggcagaa
     5221 gtatgcaaag catgcatctc aattagtcag caaccaggtg tggaaagtcc ccaggctccc
     5281 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata gtcccgcccc
     5341 taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg ccccatggct
     5401 gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag ctattccaga
     5461 agtagtgagg aggctttttt ggaggcctag gcttttgcaa agatcgatca agagacagga
     5521 tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc ggccgcttgg
     5581 gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc tgatgccgcc
     5641 gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga cctgtccggt
     5701 gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac gacgggcgtt
     5761 ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct gctattgggc
     5821 gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa agtatccatc
     5881 atggctgatg caatgcggcg gctgcatacg cttgatccg
//
