LOCUS       sequence-36041-e        5986 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 5986)
  AUTHORS   Subach OM, Patterson GH, Ting LM, Wang Y, Condeelis JS, Verkhusha VV
  TITLE     A photoswitchable orange-to-far-red fluorescent protein, PSmOrange.
  JOURNAL   Nat Methods. 2011 Jul 31;8(9):771-7. doi: 10.1038/nmeth.1664.
  PUBMED    21804536
REFERENCE   2  (bases 1 to 5986)
  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: "10.1038/nmeth"; 
            journalName: "Nat Methods"; date: "2011-07-31- 31"; volume: "8"; 
            issue: "9"; pages: "771-7"
COMMENT     Sequence Label: pKeratin-PSmOrange
FEATURES             Location/Qualifiers
     source          1..5986
                     /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      1935..1944
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1941..2651
                     /codon_start=1
                     /product="orange to far-red photoswitchable variant of the 
                     monomeric fluorescent protein mOrange (Subach et al., 
                     2011)"
                     /label=PSmOrange
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEENNMAIIKEFMRFKVRMEGTVNGHEFEIEGEGEGRPYEG
                     FQTAKLKVTKGGPLPFAWDILSPLFTYGSKAYVKHPADIPDYFKLSFPEGFKWERVMNY
                     EDGGVVTVTQDSSLQDGEFIYKVKMRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIRMRLKLKDGGHYTSEVKTTYKAKKSVQLPGAYIVGIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     2548..2571
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2774..2895
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2811..2830)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2865..2884
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2902..3357)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3039..3060)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3251..3270
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3384..3488
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3490..3847
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        3490..3681
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(3513..3533)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3698..3833
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             3882..4676
                     /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             3882..4676
                     /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(3936..3955)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4546..4565
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4864..4883)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4908..4955
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5284..5872
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5773..5792
                     /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 gctagcacca
      601 tgagcttcac cactcgctcc accttctcca ccaactaccg gtccctgggc tctgtccagg
      661 cgcccagcta cggcgcccgg ccggtcagca gcgcggccag cgtctatgca ggcgctgggg
      721 gctctggttc ccggatctcc gtgtcccgct ccaccagctt caggggcggc atggggtccg
      781 ggggcctggc caccgggata gccgggggtc tggcaggaat gggaggcatc cagaacgaga
      841 aggagaccat gcaaagcctg aacgaccgcc tggcctctta cctggacaga gtgaggagcc
      901 tggagaccga gaaccggagg ctggagagca aaatccggga gcacttggag aagaagggac
      961 cccaggtcag agactggagc cattacttca agatcatcga ggacctgagg gctcagatct
     1021 tcgcaaatac tgtggacaat gcccgcatcg ttctgcagat tgacaatgcc cgtcttgctg
     1081 ctgatgactt tagagtcaag tatgagacag agctggccat gcgccagtct gtggagaacg
     1141 acatccatgg gctccgcaag gtcattgatg acaccaatat cacacgactg cagctggaga
     1201 cagagatcga ggctctcaag gaggagctgc tcttcatgaa gaagaaccac gaagaggaag
     1261 taaaaggcct acaagcccag attgccagct ctgggttgac cgtggaggta gatgccccca
     1321 aatctcagga cctcgccaag atcatggcag acatccgggc ccaatatgac gagctggctc
     1381 ggaagaaccg agaggagcta gacaagtact ggtctcagca gattgaggag agcaccacag
     1441 tggtcaccac acagtctgct gaggttggag ctgctgagac gacgctcaca gagctgagac
     1501 gtacagtcca gtccttggag atcgacctgg actccatgag aaatctgaag gccagcttgg
     1561 agaacagcct gagggaggtg gaggcccgct acgccctaca gatggagcag ctcaacggga
     1621 tcctgctgca ccttgagtca gagctggcac agacccgggc agagggacag cgccaggccc
     1681 aggagtatga ggccctgctg aacatcaagg tcaagctgga ggctgagatc gccacctacc
     1741 gccgcctgct ggaagatggc gaggacttta atcttggtga tgccttggac agcagcaact
     1801 ccatgcaaac catccaaaag accaccaccc gccggatagt ggatggcaaa gtggtgtctg
     1861 agaccaatga caccaaagtt ctgaggcatc gaattctgca gtcgacggta ccgcgggccc
     1921 gggatccacc ggtcgccacc atggtgagca agggcgagga gaataatatg gccatcatca
     1981 aggagttcat gcgcttcaag gtgcgcatgg agggcaccgt gaacggccac gagttcgaga
     2041 tcgagggcga gggcgagggc cgcccctacg agggctttca gaccgctaag ctgaaggtga
     2101 ccaagggcgg ccccctgccc ttcgcctggg acatcctgtc ccctctcttc acctacggct
     2161 ccaaggccta cgtgaagcac cccgccgaca tccccgacta cttcaagctg tccttccccg
     2221 agggcttcaa gtgggagcgc gtgatgaact acgaggacgg cggcgtggtg accgtgaccc
     2281 aggactcctc actgcaggac ggcgagttca tctacaaggt gaagatgcgc ggcaccaact
     2341 tcccctccga cggccccgtg atgcagaaga agaccatggg ctgggaggcc tcctccgagc
     2401 ggatgtaccc cgaggacggc gccctgaagg gcgagatcag gatgaggctg aagctgaagg
     2461 acggcggcca ctacacctcc gaggtcaaga ccacctacaa ggccaagaag tccgtgcagc
     2521 tgcccggcgc ctacatcgtc ggcatcaagc tggacatcac ctcccacaac gaggactaca
     2581 ccatcgtgga acagtacgaa cgcgccgagg gccgccactc caccggcggc atggacgagc
     2641 tgtacaagta aagcggccgc gactctagat cataatcagc cataccacat ttgtagaggt
     2701 tttacttgct ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc
     2761 aattgttgtt gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     2821 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     2881 catcaatgta tcttaaggcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa
     2941 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
     3001 aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac
     3061 tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc
     3121 cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa
     3181 atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg
     3241 cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg
     3301 tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag
     3361 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     3421 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     3481 ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa
     3541 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     3601 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     3661 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     3721 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     3781 cgcctcggcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     3841 ttgcaaagat cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat
     3901 tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac
     3961 agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc
     4021 tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc
     4081 tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag
     4141 cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc
     4201 ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg
     4261 atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc
     4321 ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc
     4381 cagccgaact gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga
     4441 cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca
     4501 tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg
     4561 atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg
     4621 ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg
     4681 gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     4741 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     4801 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct
     4861 aactgaaaca cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag
     4921 acagaataaa acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     4981 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     5041 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     5101 ggcggcaggc cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact
     5161 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     5221 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     5281 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     5341 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     5401 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     5461 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     5521 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     5581 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     5641 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     5701 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     5761 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     5821 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     5881 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     5941 tgcgttatcc cctgattctg tggataaccg tattaccgcc atgcat
//
