LOCUS       sequence-42488-e        6101 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6101)
  AUTHORS   Subach OM, Entenberg D, Condeelis JS, Verkhusha VV
  TITLE     A FRET-facilitated photoswitching using an orange fluorescent 
            protein with the fast photoconversion kinetics.
  JOURNAL   J Am Chem Soc. 2012 Aug 17.
  PUBMED    22900938
REFERENCE   2  (bases 1 to 6101)
  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: "J Am Chem 
            Soc. 2012 Aug 17."
COMMENT     Sequence Label: pVimentin-PSmOrange2
FEATURES             Location/Qualifiers
     source          1..6101
                     /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"
     regulatory      2050..2059
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             2056..2763
                     /codon_start=1
                     /product="enhanced orange to far-red photoswitchable 
                     monomeric fluorescent protein (Subach et al., 2012)"
                     /label=PSmOrange2
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVHMEGTVNGHEFEIEGEGEGHPYEG
                     FQTAKLKVTKGGPLPFAWDILSPLITYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIRMRLKLKDGGHYTSEVKTTYKAKKSVLLPGAYIVGIKLDITSHNEDYTIVEQYERS
                     EARHSTGGMDELYK"
     primer_bind     2663..2686
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2889..3010
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2926..2945)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2980..2999
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3017..3472)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3154..3175)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3366..3385
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3499..3603
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3605..3962
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3605..3796
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3628..3648)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3813..3948
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3997..4791
                     /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             3997..4791
                     /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(4051..4070)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4661..4680
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4979..4998)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5023..5070
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5399..5987
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5888..5907
                     /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 gctagcgcta
      601 ccggactcag atctcgagct caagcttcga attcccatgt ccaccaggtc cgtgtcctcg
      661 tcctcctacc gcaggatgtt cggcggcccg ggcaccgcga gccggccgag ctccagccgg
      721 agctacgtga ctacgtccac ccgcacctac agcctgggca gcgcgctgcg ccccagcacc
      781 agccgcagcc tctacgcctc gtccccgggc ggcgtgtatg ccacgcgctc ctctgccgtg
      841 cgcctgcgga gcagcgtgcc cggggtgcgg ctcctgcagg actcggtgga cttctcgctg
      901 gccgacgcca tcaacaccga gttcaagaac acccgcacca acgagaaggt ggagctgcag
      961 gagctgaatg accgcttcgc caactacatc gacaaggtgc gcttcctgga gcagcagaat
     1021 aagatcctgc tggccgagct cgagcagctc aagggccaag gcaagtcgcg cctgggggac
     1081 ctctacgagg aggagatgcg ggagctgcgc cggcaggtgg accagctaac caacgacaaa
     1141 gcccgcgtcg aggtggagcg cgacaacctg gccgaggaca tcatgcgcct ccgggagaaa
     1201 ttgcaggagg agatgcttca gagagaggaa gccgaaaaca ccctgcaatc tttcagacag
     1261 gatgttgaca atgcgtctct ggcacgtctt gaccttgaac gcaaagtgga atctttgcaa
     1321 gaagagattg cctttttgaa gaaactccac gaagaggaaa tccaggagct gcaggctcag
     1381 attcaggaac agcatgtcca aatcgatgtg gatgtttcca agcctgacct cacggctgcc
     1441 ctgcgtgacg tacgtcagca atatgaaagt gtggctgcca agaacctgca ggaggcagaa
     1501 gaatggtaca aatccaagtt tgctgacctc tctgaggctg ccaaccggaa caatgacgcc
     1561 ctgcgccagg caaagcagga gtccactgag taccggagac aggtgcagtc cctcacctgt
     1621 gaagtggatg cccttaaagg aaccaatgag tccctggaac gccagatgcg tgaaatggaa
     1681 gagaactttg ccgttgaagc tgctaactac caagacacta ttggccgcct gcaggatgag
     1741 attcagaata tgaaggagga aatggctcgt caccttcgtg aataccaaga cctgctcaat
     1801 gttaagatgg cccttgacat tgagattgcc acctacagga agctgctgga aggcgaggag
     1861 agcaggattt ctctgcctct tccaaacttt tcctccctga acctgaggga aactaatctg
     1921 gattcactcc ctctggttga tacccactca aaaaggacac ttctgattaa gacggttgaa
     1981 actagagatg gacaggttat caacgaaact tctcagcatc acgatgacct tgaaggggat
     2041 ccaccggtcg ccaccatggt gagcaagggc gaggagaata atatggccat cattaaggag
     2101 ttcatgcgct tcaaggtgca catggagggc actgtgaacg gccacgagtt cgagatcgag
     2161 ggcgagggcg agggccaccc ctacgagggc tttcagaccg ctaagctgaa ggtgaccaag
     2221 ggcggccccc tgcccttcgc ctgggacatc ctgtcccctc tcatcaccta cggctccaag
     2281 gcctacgtga agcaccccgc cgacatcccc gactacttca agctgtcctt ccccgagggc
     2341 ttcaagtggg agcgcgtgat gaactacgag gacggcggcg tggtgaccgt gacccaggac
     2401 tcctctctgc aggacggcga gttcatctac aaggtgaaga tgcgcggcac caacttcccc
     2461 tccgacggcc ccgtgatgca gaagaagacc atgggctggg aggcctcctc cgagcggatg
     2521 taccccgagg acggcgccct gaagggcgag atcaggatga ggctgaagct gaaggatggc
     2581 ggccactaca cctccgaggt caagactacc tacaaggcca agaagtccgt gctgctgccc
     2641 ggcgcctaca tcgtcggcat caagctggac atcacctccc acaacgagga ctacaccatc
     2701 gtggaacagt acgaacgctc cgaggcccgc cactccaccg gcggcatgga cgagctgtac
     2761 aagtaaagcg gccgcgactc tagatcataa tcagccatac cacatttgta gaggttttac
     2821 ttgctttaaa aaacctccca cacctccccc tgaacctgaa acataaaatg aatgcaattg
     2881 ttgttgttaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     2941 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     3001 atgtatctta aggcgtaaat tgtaagcgtt aatattttgt taaaattcgc gttaaatttt
     3061 tgttaaatca gctcattttt taaccaatag gccgaaatcg gcaaaatccc ttataaatca
     3121 aaagaataga ccgagatagg gttgagtgtt gttccagttt ggaacaagag tccactatta
     3181 aagaacgtgg actccaacgt caaagggcga aaaaccgtct atcagggcga tggcccacta
     3241 cgtgaaccat caccctaatc aagttttttg gggtcgaggt gccgtaaagc actaaatcgg
     3301 aaccctaaag ggagcccccg atttagagct tgacggggaa agccggcgaa cgtggcgaga
     3361 aaggaaggga agaaagcgaa aggagcgggc gctagggcgc tggcaagtgt agcggtcacg
     3421 ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc tacagggcgc gtcaggtggc
     3481 acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat
     3541 atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag
     3601 agtcctgagg cggaaagaac cagctgtgga atgtgtgtca gttagggtgt ggaaagtccc
     3661 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccaggt
     3721 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     3781 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     3841 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     3901 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     3961 aagatcgatc aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac
     4021 gcaggttctc cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca
     4081 atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt
     4141 gtcaagaccg acctgtccgg tgccctgaat gaactgcaag acgaggcagc gcggctatcg
     4201 tggctggcca cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga
     4261 agggactggc tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct
     4321 cctgccgaga aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg
     4381 gctacctgcc cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg
     4441 gaagccggtc ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc
     4501 gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg aggatctcgt cgtgacccat
     4561 ggcgatgcct gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac
     4621 tgtggccggc tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt
     4681 gctgaagagc ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct
     4741 cccgattcgc agcgcatcgc cttctatcgc cttcttgacg agttcttctg agcgggactc
     4801 tggggttcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat ttcgattcca
     4861 ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga
     4921 tcctccagcg cggggatctc atgctggagt tcttcgccca ccctaggggg aggctaactg
     4981 aaacacggaa ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga
     5041 ataaaacgca cggtgttggg tcgtttgttc ataaacgcgg ggttcggtcc cagggctggc
     5101 actctgtcga taccccaccg agaccccatt ggggccaata cgcccgcgtt tcttcctttt
     5161 ccccacccca ccccccaagt tcgggtgaag gcccagggct cgcagccaac gtcggggcgg
     5221 caggccctgc catagcctca ggttactcat atatacttta gattgattta aaacttcatt
     5281 tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt
     5341 aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt
     5401 gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag
     5461 cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta actggcttca
     5521 gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc caccacttca
     5581 agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg
     5641 ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg
     5701 cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct
     5761 acaccgaact gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga
     5821 gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc
     5881 ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg
     5941 agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg
     6001 cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt
     6061 tatcccctga ttctgtggat aaccgtatta ccgccatgca t
//
