LOCUS       sequence-149114-        8204 bp    DNA     circular SYN 14-APR-2025
DEFINITION  high FRET control for ActTS-GR.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8204)
  AUTHORS   Yamashita S, Tsuboi T, Ishinabe N, Kitaguchi T, Michiue T
  TITLE     Wide and high resolution tension measurement using FRET in embryo.
  JOURNAL   Sci Rep. 2016 Jun 23;6:28535. doi: 10.1038/srep28535.
  PUBMED    27335157
REFERENCE   2  (bases 1 to 8204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2016-06-23"; volume: "6:28535. doi"; pages: " 
            10.1038/srep28535"
COMMENT     Sequence Label: hiActTS-GR
FEATURES             Location/Qualifiers
     source          1..8204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        61..364
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        365..568
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     519..539
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             3279..3986
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(3424..3442)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     3692..3711
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     CDS             join(4119..4121,4122..4124,4125..4835)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(4119..4121,4122..4124,4125..4835)
                     /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(4164..4185)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(4425..4444)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4772..4793
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    4992..5113
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5029..5048)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5083..5102
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(5120..5575)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5257..5278)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5469..5488
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        5602..5706
                     /gene="bla"
                     /label=AmpR promoter
     promoter        5708..6065
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        5708..5899
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(5731..5751)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      5916..6051
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             6100..6894
                     /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             6100..6894
                     /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(6154..6173)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     6764..6783
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(7082..7101)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    7126..7173
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      7502..8090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7991..8010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
       61 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      121 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      181 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      241 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      301 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      361 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      421 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      481 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      541 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagcatgg
      601 accattatga ttctcagcaa accaacgatt acatgcagcc agaagaggac tgggaccggg
      661 acctgctcct ggacccggcc tgggagaagc agcagagaaa gacattcacg gcatggtgta
      721 actcccacct ccggaaggcg gggacacaga tcgagaacat cgaagaggac ttccgggatg
      781 gcctgaagct catgctgctg ctggaggtca tctcaggtga acgcttggcc aagccagagc
      841 gaggcaagat gagagtgcac aagatctcca acgtcaacaa ggccctggat ttcatagcca
      901 gcaaaggcgt caaactggtg tccatcggag ccgaagaaat cgtggatggg aatgtgaaga
      961 tgaccctggg catgatctgg accatcatcc tgcgctttgc catccaggac atctccgtgg
     1021 aagagacttc agccaaggaa gggctgctcc tgtggtgtca gagaaagaca gccccttaca
     1081 aaaatgtcaa catccagaac ttccacataa gctggaagga tggcctcggc ttctgtgctt
     1141 tgatccaccg acaccggccc gagctgattg actacgggaa gctgcggaag gatgatccac
     1201 tcacaaatct gaatacggct tttgacgtgg cagagaagta cctggacatc cccaagatgc
     1261 tggatgccga agacatcgtt ggaactgccc gaccggatga gaaagccatc atgacttacg
     1321 tgtctagctt ctaccacgcc ttctctggag cccagaaggc ggagacagca gccaatcgca
     1381 tctgcaaggt gttggccgtc aaccaggaga acgagcagct tatggaagac tacgagaagc
     1441 tggccagtga tctgttggag tggatccgcc gcacaatccc gtggctggag aaccgggtgc
     1501 ccgagaacac catgcatgcc atgcaacaga agctggagga cttccgggac taccggcgcc
     1561 tgcacaagcc gcccaaggtg caggagaagt gccagctgga gatcaacttc aacacgctgc
     1621 agaccaagct gcggctcagc aaccggcctg ccttcatgcc ctctgagggc aggatggtct
     1681 cggacatcaa caatgcctgg ggctgcctgg agcaggtgga gaagggctat gaggagtggt
     1741 tgctgaatga gatccggagg ctggagcgac tggaccacct ggcagagaag ttccggcaga
     1801 aggcctccat ccacgaggcc tggactgacg gcaaagaggc catgctgcga cagaaggact
     1861 atgagaccgc caccctctcg gagatcaagg ccctgctcaa gaagcatgag gccttcgaga
     1921 gtgacctggc tgcccaccag gaccgtgtgg agcagattgc cgccatcgca caggagctca
     1981 atgagctgga ctattatgac tcacccagtg tcaacgcccg ttgccaaaag atctgtgacc
     2041 agtgggacaa tctgggggcc ctaactcaga agcgaaggga agctctggag cggaccgaga
     2101 aactgctgga gaccattgac cagctgtact tggagtatgc caagcgggct gcacccttca
     2161 acaactggat ggagggggcc atggaggacc tgcaggacac cttcattgtg cacaccattg
     2221 aggagatcca gggactgacc acagcccatg agcagttcaa ggccaccctc cctgatgccg
     2281 acaaggagcg cctggccatc ctgggcatcc acaatgaggt gtccaagatt gtccagacct
     2341 accacgtcaa tatggcgggc accaacccct acacaaccat cacgcctcag gagatcaatg
     2401 gcaaatggga ccacgtgcgg cagctggtgc ctcggaggga ccaagctctg acggaggagc
     2461 atgcccgaca gcagcacaat gagaggctac gcaagcagtt tggagcccag gccaatgtca
     2521 tcgggccctg gatccagacc aagatggagg agatcgggag gatctccatt gagatgcatg
     2581 ggaccctgga ggaccagctc agccacctgc ggcagtatga gaagagcatc gtcaactaca
     2641 agccaaagat tgatcagctg gagggcgacc accagctcat ccaggaggcg ctcatcttcg
     2701 acaacaagca caccaactac accatggagc acatccgtgt gggctgggag cagctgctca
     2761 ccaccatcgc caggaccatc aatgaggtag agaaccagat cctgacccgg gatgccaagg
     2821 gcatcagcca ggagcagatg aatgagttcc gggcctcctt caaccacttt gaccgggatc
     2881 actccggcac actgggtccc gaggagttca aagcctgcct catcagcttg ggttatgata
     2941 ttggcaacga cccccaggga gaagcagaat ttgcccgcat catgagcatt gtggacccca
     3001 accgcctggg ggtagtgaca ttccaggcct tcattgactt catgtcccgc gagacagccg
     3061 acacagatac agcagaccaa gtcatggctt ccttcaagat cctggctggg gacaagaact
     3121 acattaccat ggacgagctg cgccgcgagc tgccacccga ccaggctgag tactgcatcg
     3181 cgcggatggc cccctacacc ggccccgact ccgtgccagg tgctctggac tacatgtcct
     3241 tctccacggc gctgtacggc gagagtgacc tcaccggtat ggtgagcaag ggcgaggagg
     3301 ataacatggc catcatcaag gagttcatgc gcttcaaggt gcacatggag ggctccgtga
     3361 acggccacga gttcgagatc gagggcgagg gcgagggccg cccctacgag ggcacccaga
     3421 ccgccaagct gaaggtgacc aagggtggcc ccctgccctt cgcctgggac atcctgtccc
     3481 ctcagttcat gtacggctcc aaggcctacg tgaagcaccc cgccgacatc cccgactact
     3541 tgaagctgtc cttccccgag ggcttcaagt gggagcgcgt gatgaacttc gaggacggcg
     3601 gcgtggtgac cgtgacccag gactcctccc ttcaggacgg cgagttcatc tacaaggtga
     3661 agctgcgcgg caccaacttc ccctccgacg gccccgtaat gcagaagaag accatgggct
     3721 gggaggcctc ctccgagcgg atgtaccccg aggacggcgc cctgaagggc gagatcaagc
     3781 agaggctgaa gctgaaggac ggcggccact acgacgctga ggtcaagacc acctacaagg
     3841 ccaagaagcc cgtgcagctg cccggcgcct acaacgtcaa catcaagttg gacatcacct
     3901 cccacaacga ggactacacc atcgtggaac agtacgaacg cgccgagggc cgccactcca
     3961 ccggcggcat ggacgagctg tacaaggaat tcgggccagg tggtgcaggg ccaggtggtg
     4021 cagggccagg tggtgcaggg ccaggtggtg cagggcccgg tggtgcaggc ccaggtggtg
     4081 caggtccagg tggtgcaggt ccaggtggtg ctggatccat ggtgagcaag ggcgaggagc
     4141 tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac ggccacaagt
     4201 tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc ctgaagttca
     4261 tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc ctgacctacg
     4321 gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc ttcaagtccg
     4381 ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac ggcaactaca
     4441 agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc gagctgaagg
     4501 gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac aactacaaca
     4561 gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg aacttcaaga
     4621 tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag cagaacaccc
     4681 ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc cagtccgccc
     4741 tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc gtgaccgccg
     4801 ccgggatcac tctcggcatg gacgagctgt acaagagcgg ccgctaaggt accgcgggcc
     4861 cgggatccac cggatctaga taactgatca taatcagcca taccacattt gtagaggttt
     4921 tacttgcttt aaaaaacctc ccacacctcc ccctgaacct gaaacataaa atgaatgcaa
     4981 ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5041 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5101 tcaatgtatc ttaacgcgta aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     5161 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5221 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5281 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5341 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5401 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5461 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5521 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtcaggt
     5581 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     5641 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     5701 aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg tcagttaggg tgtggaaagt
     5761 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     5821 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     5881 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     5941 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     6001 cctcggcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     6061 gcaaagatcg atcaagagac aggatgagga tcgtttcgca tgattgaaca agatggattg
     6121 cacgcaggtt ctccggccgc ttgggtggag aggctattcg gctatgactg ggcacaacag
     6181 acaatcggct gctctgatgc cgccgtgttc cggctgtcag cgcaggggcg cccggttctt
     6241 tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc aagacgaggc agcgcggcta
     6301 tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc tcgacgttgt cactgaagcg
     6361 ggaagggact ggctgctatt gggcgaagtg ccggggcagg atctcctgtc atctcacctt
     6421 gctcctgccg agaaagtatc catcatggct gatgcaatgc ggcggctgca tacgcttgat
     6481 ccggctacct gcccattcga ccaccaagcg aaacatcgca tcgagcgagc acgtactcgg
     6541 atggaagccg gtcttgtcga tcaggatgat ctggacgaag agcatcaggg gctcgcgcca
     6601 gccgaactgt tcgccaggct caaggcgagc atgcccgacg gcgaggatct cgtcgtgacc
     6661 catggcgatg cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc
     6721 gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat
     6781 attgctgaag agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc
     6841 gctcccgatt cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgagcggga
     6901 ctctggggtt cgaaatgacc gaccaagcga cgcccaacct gccatcacga gatttcgatt
     6961 ccaccgccgc cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga
     7021 tgatcctcca gcgcggggat ctcatgctgg agttcttcgc ccaccctagg gggaggctaa
     7081 ctgaaacacg gaaggagaca ataccggaag gaacccgcgc tatgacggca ataaaaagac
     7141 agaataaaac gcacggtgtt gggtcgtttg ttcataaacg cggggttcgg tcccagggct
     7201 ggcactctgt cgatacccca ccgagacccc attggggcca atacgcccgc gtttcttcct
     7261 tttccccacc ccacccccca agttcgggtg aaggcccagg gctcgcagcc aacgtcgggg
     7321 cggcaggccc tgccatagcc tcaggttact catatatact ttagattgat ttaaaacttc
     7381 atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc
     7441 cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt
     7501 cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac
     7561 cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct
     7621 tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact
     7681 tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg
     7741 ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata
     7801 aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga
     7861 cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag
     7921 ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg
     7981 agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac
     8041 ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca
     8101 acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg
     8161 cgttatcccc tgattctgtg gataaccgta ttaccgccat gcat
//
