LOCUS       Exported                6042 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pPSmOrange-Tubulin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6042)
  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 6042)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6042
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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      601..610
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             607..1314
                     /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     1214..1237
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    2830..2951
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2867..2886)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2921..2940
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2958..3413)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3095..3116)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3307..3326
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3440..3544
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3546..3903
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3569..3589)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3754..3889
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3816..3835
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3938..4732
                     /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(3992..4011)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4602..4621
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4920..4939)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4964..5011
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5340..5928
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5829..5848
                     /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 gctagcggtc
      601 gccaccatgg tgagcaaggg cgaggagaat aatatggcca tcatcaagga gttcatgcgc
      661 ttcaaggtgc gcatggaggg caccgtgaac ggccacgagt tcgagatcga gggcgagggc
      721 gagggccgcc cctacgaggg ctttcagacc gctaagctga aggtgaccaa gggcggcccc
      781 ctgcccttcg cctgggacat cctgtcccct ctcttcacct acggctccaa ggcctacgtg
      841 aagcaccccg ccgacatccc cgactacttc aagctgtcct tccccgaggg cttcaagtgg
      901 gagcgcgtga tgaactacga ggacggcggc gtggtgaccg tgacccagga ctcctcactg
      961 caggacggcg agttcatcta caaggtgaag atgcgcggca ccaacttccc ctccgacggc
     1021 cccgtgatgc agaagaagac catgggctgg gaggcctcct ccgagcggat gtaccccgag
     1081 gacggcgccc tgaagggcga gatcaggatg aggctgaagc tgaaggacgg cggccactac
     1141 acctccgagg tcaagaccac ctacaaggcc aagaagtccg tgcagctgcc cggcgcctac
     1201 atcgtcggca tcaagctgga catcacctcc cacaacgagg actacaccat cgtggaacag
     1261 tacgaacgcg ccgagggccg ccactccacc ggcggcatgg acgagctgta caagggtgga
     1321 ggaggttccg gactcagatc tcgagtgcgt gagtgcatct ccatccacgt tggccaggct
     1381 ggtgtccaga ttggcaatgc ctgctgggag ctctactgcc tggaacacgg catccagccc
     1441 gatggccaga tgccaagtga caagaccatt gggggaggag atgactcctt caacaccttc
     1501 ttcagtgaga cgggcgctgg caagcacgtg ccccgggctg tgtttgtaga cttggaaccc
     1561 acagtcattg atgaagttcg cactggcacc taccgccagc tcttccaccc tgagcagctc
     1621 atcacaggca aggaagatgc tgccaataac tatgcccgag ggcactacac cattggcaag
     1681 gagatcattg accttgtgtt ggaccgaatt cgcaagctgg ctgaccagtg caccggtctt
     1741 cagggcttct tggttttcca cagctttggt gggggaactg gttctgggtt cacctccctg
     1801 ctcatggaac gtctctcagt tgattatggc aagaagtcca agctggagtt ctccatttac
     1861 ccagcacccc aggtttccac agctgtagtt gagccctaca actccatcct caccacccac
     1921 accaccctgg agcactctga ttgtgccttc atggtagaca atgaggccat ctatgacatc
     1981 tgtcgtagaa acctcgatat cgagcgccca acctacacta accttaaccg ccttattagc
     2041 cagattgtgt cctccatcac tgcttccctg agatttgatg gagccctgaa tgttgacctg
     2101 acagaattcc agaccaacct ggtgccctac ccccgcatcc acttccctct ggccacatat
     2161 gcccctgtca tctctgctga gaaagcctac catgaacagc tttctgtagc agagatcacc
     2221 aatgcttgct ttgagccagc caaccagatg gtgaaatgtg accctcgcca tggtaaatac
     2281 atggcttgct gcctgttgta ccgtggtgac gtggttccca aagatgtcaa tgctgccatt
     2341 gccaccatca aaaccaagcg cagcatccag tttgtggatt ggtgccccac tggcttcaag
     2401 gttggcatca actaccagcc tcccactgtg gtgcctggtg gagacctggc caaggtacag
     2461 agagctgtgt gcatgctgag caacaccaca gccattgctg aggcctgggc tcgcctggac
     2521 cacaagtttg acctgatgta tgccaagcgt gcctttgttc actggtacgt gggtgagggg
     2581 atggaggaag gcgagttttc agaggcccgt gaagatatgg ctgcccttga gaaggattat
     2641 gaggaggttg gtgtggattc tgttgaagga gagggtgagg aagaaggaga ggaatactaa
     2701 ggatccaccg gatctagata actgatcata atcagccata ccacatttgt agaggtttta
     2761 cttgctttaa aaaacctccc acacctcccc ctgaacctga aacataaaat gaatgcaatt
     2821 gttgttgtta acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     2881 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     2941 aatgtatctt aacgcgtaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     3001 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     3061 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     3121 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     3181 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     3241 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     3301 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     3361 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtcaggtgg
     3421 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3481 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3541 gagtcctgag gcggaaagaa ccagctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     3601 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     3661 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     3721 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     3781 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     3841 tcggcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     3901 aaagatcgat caagagacag gatgaggatc gtttcgcatg attgaacaag atggattgca
     3961 cgcaggttct ccggccgctt gggtggagag gctattcggc tatgactggg cacaacagac
     4021 aatcggctgc tctgatgccg ccgtgttccg gctgtcagcg caggggcgcc cggttctttt
     4081 tgtcaagacc gacctgtccg gtgccctgaa tgaactgcaa gacgaggcag cgcggctatc
     4141 gtggctggcc acgacgggcg ttccttgcgc agctgtgctc gacgttgtca ctgaagcggg
     4201 aagggactgg ctgctattgg gcgaagtgcc ggggcaggat ctcctgtcat ctcaccttgc
     4261 tcctgccgag aaagtatcca tcatggctga tgcaatgcgg cggctgcata cgcttgatcc
     4321 ggctacctgc ccattcgacc accaagcgaa acatcgcatc gagcgagcac gtactcggat
     4381 ggaagccggt cttgtcgatc aggatgatct ggacgaagag catcaggggc tcgcgccagc
     4441 cgaactgttc gccaggctca aggcgagcat gcccgacggc gaggatctcg tcgtgaccca
     4501 tggcgatgcc tgcttgccga atatcatggt ggaaaatggc cgcttttctg gattcatcga
     4561 ctgtggccgg ctgggtgtgg cggaccgcta tcaggacata gcgttggcta cccgtgatat
     4621 tgctgaagag cttggcggcg aatgggctga ccgcttcctc gtgctttacg gtatcgccgc
     4681 tcccgattcg cagcgcatcg ccttctatcg ccttcttgac gagttcttct gagcgggact
     4741 ctggggttcg aaatgaccga ccaagcgacg cccaacctgc catcacgaga tttcgattcc
     4801 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     4861 atcctccagc gcggggatct catgctggag ttcttcgccc accctagggg gaggctaact
     4921 gaaacacgga aggagacaat accggaagga acccgcgcta tgacggcaat aaaaagacag
     4981 aataaaacgc acggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg
     5041 cactctgtcg ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt
     5101 tccccacccc accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg
     5161 gcaggccctg ccatagcctc aggttactca tatatacttt agattgattt aaaacttcat
     5221 ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct
     5281 taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct
     5341 tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca
     5401 gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc
     5461 agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc
     5521 aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct
     5581 gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag
     5641 gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc
     5701 tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg
     5761 agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag
     5821 cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt
     5881 gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac
     5941 gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg
     6001 ttatcccctg attctgtgga taaccgtatt accgccatgc at
//
