LOCUS       Exported                6036 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pPSmOrange2-Tubulin
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6036)
  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 6036)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6036
                     /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      607..616
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             613..1320
                     /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     1026..1045
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2824..2945
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2861..2880)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2915..2934
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2952..3407)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3089..3110)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3301..3320
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3434..3538
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3540..3897
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3563..3583)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3748..3883
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3810..3829
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3932..4726
                     /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(3986..4005)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4596..4615
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4914..4933)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4958..5005
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5334..5922
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5823..5842
                     /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 ccggtcgcca ccatggtgag caagggcgag gagaataata tggccatcat taaggagttc
      661 atgcgcttca aggtgcacat ggagggcact gtgaacggcc acgagttcga gatcgagggc
      721 gagggcgagg gccaccccta cgagggcttt cagaccgcta agctgaaggt gaccaagggt
      781 ggccccctgc ccttcgcctg ggacatcctg tcccctctca tcacctacgg ctccaaggcg
      841 tacgtgaagc accccgccga catccccgac tacttcaagc tgtccttccc cgagggcttc
      901 aagtgggagc gcgtgatgaa ctacgaggac ggcggcgtgg tgaccgtgac ccaggactcc
      961 tctctgcagg acggcgagtt catctacaag gtgaagatgc gcggcaccaa cttcccctcc
     1021 gacggccccg taatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac
     1081 cccgaggacg gcgccctgaa gggcgagatc aggatgaggc tgaagctgaa ggatggcggc
     1141 cactacacct ccgaggtcaa gactacctac aaggccaaga agtccgtgct gctgcccggc
     1201 gcctacatcg tcggcatcaa gttggacatt acctcacaca acgaggacta caccatcgtg
     1261 gaacagtatg aacgcagcga ggcccgccac tccaccggcg gcatggacga gctgtacaag
     1321 tccggactca gatctcgagt gcgtgagtgc atctccatcc acgttggcca ggctggtgtc
     1381 cagattggca atgcctgctg ggagctctac tgcctggaac acggcatcca gcccgatggc
     1441 cagatgccaa gtgacaagac cattggggga ggagatgact ccttcaacac cttcttcagt
     1501 gagacgggcg ctggcaagca cgtgccccgg gctgtgtttg tagacttgga acccacagtc
     1561 attgatgaag ttcgcactgg cacctaccgc cagctcttcc accctgagca gctcatcaca
     1621 ggcaaggaag atgctgccaa taactatgcc cgagggcact acaccattgg caaggagatc
     1681 attgaccttg tgttggaccg aattcgcaag ctggctgacc agtgcaccgg tcttcagggc
     1741 ttcttggttt tccacagctt tggtggggga actggttctg ggttcacctc cctgctcatg
     1801 gaacgtctct cagttgatta tggcaagaag tccaagctgg agttctccat ttacccagca
     1861 ccccaggttt ccacagctgt agttgagccc tacaactcca tcctcaccac ccacaccacc
     1921 ctggagcact ctgattgtgc cttcatggta gacaatgagg ccatctatga catctgtcgt
     1981 agaaacctcg atatcgagcg cccaacctac actaacctta accgccttat tagccagatt
     2041 gtgtcctcca tcactgcttc cctgagattt gatggagccc tgaatgttga cctgacagaa
     2101 ttccagacca acctggtgcc ctacccccgc atccacttcc ctctggccac atatgcccct
     2161 gtcatctctg ctgagaaagc ctaccatgaa cagctttctg tagcagagat caccaatgct
     2221 tgctttgagc cagccaacca gatggtgaaa tgtgaccctc gccatggtaa atacatggct
     2281 tgctgcctgt tgtaccgtgg tgacgtggtt cccaaagatg tcaatgctgc cattgccacc
     2341 atcaaaacca agcgcagcat ccagtttgtg gattggtgcc ccactggctt caaggttggc
     2401 atcaactacc agcctcccac tgtggtgcct ggtggagacc tggccaaggt acagagagct
     2461 gtgtgcatgc tgagcaacac cacagccatt gctgaggcct gggctcgcct ggaccacaag
     2521 tttgacctga tgtatgccaa gcgtgccttt gttcactggt acgtgggtga ggggatggag
     2581 gaaggcgagt tttcagaggc ccgtgaagat atggctgccc ttgagaagga ttatgaggag
     2641 gttggtgtgg attctgttga aggagagggt gaggaagaag gagaggaata ctaaggatcc
     2701 accggatcta gataactgat cataatcagc cataccacat ttgtagaggt tttacttgct
     2761 ttaaaaaacc tcccacacct ccccctgaac ctgaaacata aaatgaatgc aattgttgtt
     2821 gttaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     2881 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     2941 tcttaacgcg taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     3001 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     3061 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     3121 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     3181 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     3241 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     3301 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     3361 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcag gtggcacttt
     3421 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3481 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtcc
     3541 tgaggcggaa agaaccagct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     3601 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     3661 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     3721 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     3781 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctcggcc
     3841 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaagat
     3901 cgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     3961 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4021 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4081 gaccgacctg tccggtgccc tgaatgaact gcaagacgag gcagcgcggc tatcgtggct
     4141 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4201 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4261 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4321 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4381 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4441 gttcgccagg ctcaaggcga gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4501 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4561 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4621 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4681 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4741 ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc
     4801 gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc
     4861 cagcgcgggg atctcatgct ggagttcttc gcccacccta gggggaggct aactgaaaca
     4921 cggaaggaga caataccgga aggaacccgc gctatgacgg caataaaaag acagaataaa
     4981 acgcacggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     5041 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     5101 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     5161 cctgccatag cctcaggtta ctcatatata ctttagattg atttaaaact tcatttttaa
     5221 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     5281 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     5341 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     5401 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     5461 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     5521 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     5581 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     5641 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     5701 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     5761 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     5821 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     5881 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     5941 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     6001 cctgattctg tggataaccg tattaccgcc atgcat
//
