LOCUS       sequence-217987-        6601 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Force free control for Actin-cpstFRET-Actin.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6601)
  AUTHORS   Guo J, Wang Y, Sachs F, Meng F
  TITLE     Actin stress in cell reprogramming.
  JOURNAL   Proc Natl Acad Sci U S A. 2014 Dec 9;111(49):E5252-61. doi: 
            10.1073/pnas.1411683111. Epub 2014 Nov 24.
  PUBMED    25422450
REFERENCE   2  (bases 1 to 6601)
  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: 
            "10.1073/pnas.1411683111"; journalName: "Proc Natl Acad Sci U S A"; 
            date: "2014-12-9- 9"; volume: "111"; issue: "49"; pages: "E5252-61"
COMMENT     Sequence Label: cpstFRET-Actin
FEATURES             Location/Qualifiers
     source          1..6601
                     /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             615..1349
                     /codon_start=1
                     /product="circularly permuted form of the Venus yellow 
                     fluorescent protein (Nagai et al., 2004)"
                     /label=cp173Venus
                     /note="efficient FRET acceptor for CFP"
                     /translation="MGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSKLSKDPNEKRDH
                     MVLLEFVTAAGITLGMDELYKEFGGGGGVSKGEELFTGVVPILVELDGDVNGHKFSVSG
                     EGEGDATYGKLTLKLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPE
                     GYVQERTIFFKDDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHN
                     VYITADKQKNGIKANFKILHNIE"
     CDS             join(1584..1586,1587..2099)
                     /codon_start=1
                     /product="N-terminal fragment of mVenus for use in 
                     bimolecular fluorescence complementation (BiFC) (Kodama and
                     Hu, 2010)"
                     /label=VN173
                     /translation="VSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTWGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNAISDNVYITADKQKNGIKA
                     NFKIRHNIE"
     polyA_signal    3389..3510
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3426..3445)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3480..3499
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3517..3972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3654..3675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3866..3885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3999..4103
                     /gene="bla"
                     /label=AmpR promoter
     promoter        4105..4462
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        4105..4296
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(4128..4148)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4313..4448
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             4497..5291
                     /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             4497..5291
                     /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(4551..4570)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5161..5180
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5479..5498)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5523..5570
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5899..6487
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6388..6407
                     /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 gctagccagg
      601 gacccggggt accaatgggc ggcgtgcagc tcgccgacca ctaccagcag aacaccccca
      661 tcggcgacgg ccccgtgctg ctgcccgaca accactacct gagctaccag tccaaactga
      721 gcaaagaccc caacgagaag cgcgatcaca tggtcctgct ggagttcgtg accgccgccg
      781 ggatcactct cggcatggac gagctgtaca aggaattcgg cggcggtggc ggggtgagca
      841 agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa
      901 acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac ggcaagctga
      961 ccctgaagct gatctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca
     1021 ccctgggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag cagcacgact
     1081 tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg
     1141 acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca
     1201 tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt
     1261 acaactacaa cagccacaac gtctatatca ccgccgacaa gcagaagaac ggcatcaagg
     1321 ccaacttcaa gatcctccac aacatcgagg acggcggcag atctatgggc agcgtgcagc
     1381 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     1441 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     1501 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     1561 aggaattcgg cggcggtggc ggggtgagca agggcgagga gctgttcacc ggggtggtgc
     1621 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     1681 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     1741 tgcccgtgcc ctggcccacc ctcgtgacca ccctgacctg gggcgtgcag tgcttcgccc
     1801 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     1861 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     1921 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     1981 acggcaacat cctggggcac aagctggagt acaacgccat cagcgacaac gtctatatca
     2041 ccgccgacaa gcagaagaac ggcatcaagg ccaacttcaa gatccgccac aacatcgagg
     2101 acgagctcgg tgggggagga ggtatggatg atgatatcgc cgcgctcgtc gtcgacaacg
     2161 gctccggcat gtgcaaggcc ggcttcgcgg gcgacgatgc cccccgggcc gtcttcccct
     2221 ccatcgtggg gcgccccagg caccagggcg tgatggtggg catgggtcag aaggattcct
     2281 atgtgggcga cgaggcccag agcaagagag gcatcctcac cctgaagtac cccatcgagc
     2341 acggcatcgt caccaactgg gacgacatgg agaaaatctg gcaccacacc ttctacaatg
     2401 agctgcgtgt ggctcccgag gagcaccccg tgctgctgac cgaggccccc ctgaacccca
     2461 aggccaaccg cgagaagatg acccagatca tgtttgagac cttcaacacc ccagccatgt
     2521 acgttgctat ccaggctgtg ctatccctgt acgcctctgg ccgtaccact ggcatcgtga
     2581 tggactccgg tgacggggtc acccacactg tgcccatcta cgaggggtat gccctccccc
     2641 atgccatcct gcgtctggac ctggctggcc gggacctgac tgactacctc atgaagatcc
     2701 tcaccgagcg cggctacagc ttcaccacca cggccgagcg ggaaatcgtg cgtgacatta
     2761 aggagaagct gtgctacgtc gccctggact tcgagcaaga gatggccacg gctgcttcca
     2821 gctcctccct ggagaagagc tacgagctgc ctgacggcca ggtcatcacc attggcaatg
     2881 agcggttccg ctgccctgag gcactcttcc agccttcctt cctgggcatg gagtcctgtg
     2941 gcatccacga aactaccttc aactccatca tgaagtgtga cgtggacatc cgcaaagacc
     3001 tgtacgccaa cacagtgctg tctggcggca ccaccatgta ccctggcatt gccgacagga
     3061 tgcagaagga gatcactgcc ctggcaccca gcacaatgaa gatcaagatc attgctcctc
     3121 ctgagcgcaa gtactccgtg tggatcggcg gctccatcct ggcctcgctg tccaccttcc
     3181 agcagatgtg gatcagcaag caggagtatg acgagtccgg cccctccatc gtccaccgca
     3241 aatgcttcta aagggcccgg gatccaccgg atctagataa ctgatcataa tcagccatac
     3301 cacatttgta gaggttttac ttgctttaaa aaacctccca cacctccccc tgaacctgaa
     3361 acataaaatg aatgcaattg ttgttgttaa cttgtttatt gcagcttata atggttacaa
     3421 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     3481 tggtttgtcc aaactcatca atgtatctta aggcgtaaat tgtaagcgtt aatattttgt
     3541 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3601 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3661 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3721 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3781 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3841 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3901 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3961 tacagggcgc gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     4021 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     4081 aataatattg aaaaaggaag agtcctgagg cggaaagaac cagctgtgga atgtgtgtca
     4141 gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     4201 caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4261 aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc
     4321 cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt tttttattta
     4381 tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag gaggcttttt
     4441 tggaggccta ggcttttgca aagatcgatc aagagacagg atgaggatcg tttcgcatga
     4501 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     4561 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     4621 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     4681 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     4741 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     4801 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     4861 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     4921 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     4981 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     5041 aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc
     5101 gcttttctgg attcatcgac tgtggccggc tgggtgtggc ggaccgctat caggacatag
     5161 cgttggctac ccgtgatatt gctgaagagc ttggcggcga atgggctgac cgcttcctcg
     5221 tgctttacgg tatcgccgct cccgattcgc agcgcatcgc cttctatcgc cttcttgacg
     5281 agttcttctg agcgggactc tggggttcga aatgaccgac caagcgacgc ccaacctgcc
     5341 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     5401 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     5461 ccctaggggg aggctaactg aaacacggaa ggagacaata ccggaaggaa cccgcgctat
     5521 gacggcaata aaaagacaga ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg
     5581 ggttcggtcc cagggctggc actctgtcga taccccaccg agaccccatt ggggccaata
     5641 cgcccgcgtt tcttcctttt ccccacccca ccccccaagt tcgggtgaag gcccagggct
     5701 cgcagccaac gtcggggcgg caggccctgc catagcctca ggttactcat atatacttta
     5761 gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa
     5821 tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga
     5881 aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac
     5941 aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt
     6001 tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc
     6061 gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat
     6121 cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag
     6181 acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc
     6241 cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc tatgagaaag
     6301 cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac
     6361 aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg
     6421 gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct
     6481 atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc
     6541 tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgccatgca
     6601 t
//
