LOCUS       sequence-36057-e        5095 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 5095)
  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 5095)
  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: pH2B-PSmOrange
FEATURES             Location/Qualifiers
     source          1..5095
                     /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      1044..1053
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1050..1760
                     /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     1657..1680
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    1883..2004
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1920..1939)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1974..1993
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2011..2466)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2148..2169)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2360..2379
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2493..2597
                     /gene="bla"
                     /label=AmpR promoter
     promoter        2599..2956
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     enhancer        2599..2790
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     complement(2622..2642)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      2807..2942
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     CDS             2991..3785
                     /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             2991..3785
                     /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(3045..3064)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     3655..3674
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(3973..3992)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4017..4064
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4393..4981
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4882..4901
                     /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 attctgcagt cgacggtacc gccaccatgc
      661 cagagccagc gaagtctgct cccgccccga aaaagggctc caagaaggcg gtgactaagg
      721 cgcagaagaa agacggcaag agcgcagcgc accgcaagga gagctattcc atctatgtgt
      781 acaaggttct gaagcaggtc caccctgaca ccggcatttc gtccaaggcc atgggcatca
      841 tgaattcgtt tgtgaacgac attttcgagc gcatcgcagg tgaggcttcc cgccttgcgc
      901 attacaacaa gcgctcgacc atcacctcca gggagatcca gacggccgtg cgcctgctgc
      961 tgcctgggga gttggccaag cacgccgtgt ccgagggtac taaggccgtc accaagtaca
     1021 ccagcgctaa ggatccaccg gtcgccacca tggtgagcaa gggcgaggag aataatatgg
     1081 ccatcatcaa ggagttcatg cgcttcaagg tgcgcatgga gggcaccgtg aacggccacg
     1141 agttcgagat cgagggcgag ggcgagggcc gcccctacga gggctttcag accgctaagc
     1201 tgaaggtgac caagggcggc cccctgccct tcgcctggga catcctgtcc cctctcttca
     1261 cctacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac ttcaagctgt
     1321 ccttccccga gggcttcaag tgggagcgcg tgatgaacta cgaggacggc ggcgtggtga
     1381 ccgtgaccca ggactcctca ctgcaggacg gcgagttcat ctacaaggtg aagatgcgcg
     1441 gcaccaactt cccctccgac ggccccgtga tgcagaagaa gaccatgggc tgggaggcct
     1501 cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcagg atgaggctga
     1561 agctgaagga cggcggccac tacacctccg aggtcaagac cacctacaag gccaagaagt
     1621 ccgtgcagct gcccggcgcc tacatcgtcg gcatcaagct ggacatcacc tcccacaacg
     1681 aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc accggcggca
     1741 tggacgagct gtacaagtaa agcggccgcg actctagatc ataatcagcc ataccacatt
     1801 tgtagaggtt ttacttgctt taaaaaacct cccacacctc cccctgaacc tgaaacataa
     1861 aatgaatgca attgttgttg ttaacttgtt tattgcagct tataatggtt acaaataaag
     1921 caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt
     1981 gtccaaactc atcaatgtat cttaaggcgt aaattgtaag cgttaatatt ttgttaaaat
     2041 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     2101 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     2161 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     2221 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     2281 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     2341 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     2401 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     2461 gcgcgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     2521 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     2581 attgaaaaag gaagagtcct gaggcggaaa gaaccagctg tggaatgtgt gtcagttagg
     2641 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     2701 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     2761 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     2821 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     2881 ggccgaggcc gcctcggcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     2941 cctaggcttt tgcaaagatc gatcaagaga caggatgagg atcgtttcgc atgattgaac
     3001 aagatggatt gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact
     3061 gggcacaaca gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc
     3121 gcccggttct ttttgtcaag accgacctgt ccggtgccct gaatgaactg caagacgagg
     3181 cagcgcggct atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg
     3241 tcactgaagc gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt
     3301 catctcacct tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc
     3361 atacgcttga tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag
     3421 cacgtactcg gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg
     3481 ggctcgcgcc agccgaactg ttcgccaggc tcaaggcgag catgcccgac ggcgaggatc
     3541 tcgtcgtgac ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt
     3601 ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg
     3661 ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt
     3721 acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct
     3781 tctgagcggg actctggggt tcgaaatgac cgaccaagcg acgcccaacc tgccatcacg
     3841 agatttcgat tccaccgccg ccttctatga aaggttgggc ttcggaatcg ttttccggga
     3901 cgccggctgg atgatcctcc agcgcgggga tctcatgctg gagttcttcg cccaccctag
     3961 ggggaggcta actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc
     4021 aataaaaaga cagaataaaa cgcacggtgt tgggtcgttt gttcataaac gcggggttcg
     4081 gtcccagggc tggcactctg tcgatacccc accgagaccc cattggggcc aatacgcccg
     4141 cgtttcttcc ttttccccac cccacccccc aagttcgggt gaaggcccag ggctcgcagc
     4201 caacgtcggg gcggcaggcc ctgccatagc ctcaggttac tcatatatac tttagattga
     4261 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     4321 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     4381 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     4441 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     4501 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     4561 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     4621 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     4681 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     4741 ggagcgaacg acctacaccg aactgagata cctacagcgt gagctatgag aaagcgccac
     4801 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     4861 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     4921 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     4981 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     5041 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcca tgcat
//
