LOCUS       Exported                6388 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pPaxillin-PSmOrange
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6388)
  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 6388)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6388
                     /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"
     misc_feature    2277..2329
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      2337..2346
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2343..3053
                     /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     2950..2973
                     /label=DsRed1-C
                     /note="DsRed1, forward primer"
     polyA_signal    3176..3297
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3213..3232)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3267..3286
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(3304..3759)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3441..3462)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3653..3672
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3786..3890
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3892..4249
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3915..3935)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      4100..4235
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4162..4181
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4284..5078
                     /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(4338..4357)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4948..4967
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(5266..5285)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    5310..5357
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5686..6274
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6175..6194
                     /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 tggacgacct cgatgcctta ctggcagacc tggaatccac cacctcgcat atctctaagc
      661 gaccggtgtt tctaacagag gaaacgcctt actcctaccc aactggaaac catacgtacc
      721 aggagattgc tgtgccaccg ccggtgcccc cgccgccttc cagtgaggcc ctaaatggca
      781 ctgtgattga ccccttagac cagtggcaac ccagcgtgtc cagatacggc caccagcaac
      841 ctcagtccca gtctccgata tacagctcta gtgccaaaag ctccagtgcc tctgttccta
      901 gagacgggct cagctcccct tctccccgtg ccagtgagga ggaacacgtg tacagcttcc
      961 caaacaagca gaagtctgca gaaccatctc ccacaatgac cagcacctcc ctgggcagca
     1021 acctctcgga actggacagg cttctcctgg aactgaatgc tgttcaacat aatcccccca
     1081 gtggcttctc agcagatgaa gtcagcagaa gcccatcact gcccaatgtg accggacctc
     1141 actatgtcat cccagagagc agcagctctg caggagggaa ggctgcaccc cctacaaaag
     1201 aaaaaccaaa gcggaatggt gggcgtggga ttgaagatgt gcgtcccagt gtggagagcc
     1261 tgctggatga gctggagagc tctgtgccaa gcccagtccc tgcaatcact gtgagccaag
     1321 gggaggtgag cagcccccag cgtgtcaacg ccagtcagca gcagacccgt atctctgctt
     1381 cttcagctac acgagaactg gatgagctga tggcgtccct ctctgacttt aagttcatgg
     1441 cacaggggaa agccggcggc agcagctccc caccatccac caccccaaag cctggcagtc
     1501 agctggacac catgctggga agtctgcagt ctgacctgaa caaactgggg gtagcgacgg
     1561 tcgccaaggg tgtgtgcgga gcctgcaaga agccgattgc tgggcaggta gttacagcca
     1621 tggggaagac ctggcaccct gagcacttcg tctgcaccca ctgccaggag gagattggat
     1681 cacggaactt ctttgagcgg gatgggcagc cctactgtga gaaggactat cacaacctct
     1741 tctctcctcg ctgctactac tgcaatgggc caatccttga taaagtggtg acggctttgg
     1801 acaggacgtg gcaccctgaa cactttttct gtgcccagtg tggagttttc tttggacctg
     1861 aaggatttca tgagaaggat ggcaaagcct attgccgcaa ggactacttt gacatgtttg
     1921 ctcccaagtg tggaggctgt gcccgggcta tcctggaaaa ttacatttct gccttaaaca
     1981 ccctgtggca ccctgagtgc tttgtctgtc gggaatgttt cacaccgttc atcaacggca
     2041 gcttcttcga gcacgacggc cagccctact gcgaggtgca ttaccacgag cgccgcggct
     2101 cgctgtgctc tggctgccag aagcccatca cgggacgctg catcactgcc atgggcaaga
     2161 aatttcaccc cgaacacttt gtctgtgcct tctgcctcaa gcagctcaac aaaggaacct
     2221 tcaaagaaca gaacgacaag ccctactgtc agaactgctt tctcaaactc ttctgtctcg
     2281 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
     2341 ccatggtgag caagggcgag gagaataata tggccatcat caaggagttc atgcgcttca
     2401 aggtgcgcat ggagggcacc gtgaacggcc acgagttcga gatcgagggc gagggcgagg
     2461 gccgccccta cgagggcttt cagaccgcta agctgaaggt gaccaagggc ggccccctgc
     2521 ccttcgcctg ggacatcctg tcccctctct tcacctacgg ctccaaggcc tacgtgaagc
     2581 accccgccga catccccgac tacttcaagc tgtccttccc cgagggcttc aagtgggagc
     2641 gcgtgatgaa ctacgaggac ggcggcgtgg tgaccgtgac ccaggactcc tcactgcagg
     2701 acggcgagtt catctacaag gtgaagatgc gcggcaccaa cttcccctcc gacggccccg
     2761 tgatgcagaa gaagaccatg ggctgggagg cctcctccga gcggatgtac cccgaggacg
     2821 gcgccctgaa gggcgagatc aggatgaggc tgaagctgaa ggacggcggc cactacacct
     2881 ccgaggtcaa gaccacctac aaggccaaga agtccgtgca gctgcccggc gcctacatcg
     2941 tcggcatcaa gctggacatc acctcccaca acgaggacta caccatcgtg gaacagtacg
     3001 aacgcgccga gggccgccac tccaccggcg gcatggacga gctgtacaag taaagcggcc
     3061 gcgactctag atcataatca gccataccac atttgtagag gttttacttg ctttaaaaaa
     3121 cctcccacac ctccccctga acctgaaaca taaaatgaat gcaattgttg ttgttaactt
     3181 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     3241 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg
     3301 cgtaaattgt aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct
     3361 cattttttaa ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg
     3421 agatagggtt gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact
     3481 ccaacgtcaa agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac
     3541 cctaatcaag ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga
     3601 gcccccgatt tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga
     3661 aagcgaaagg agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca
     3721 ccacacccgc cgcgcttaat gcgccgctac agggcgcgtc aggtggcact tttcggggaa
     3781 atgtgcgcgg aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     3841 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagagt cctgaggcgg
     3901 aaagaaccag ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     3961 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4021 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4081 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4141 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctcgg cctctgagct
     4201 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaag atcgatcaag
     4261 agacaggatg aggatcgttt cgcatgattg aacaagatgg attgcacgca ggttctccgg
     4321 ccgcttgggt ggagaggcta ttcggctatg actgggcaca acagacaatc ggctgctctg
     4381 atgccgccgt gttccggctg tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc
     4441 tgtccggtgc cctgaatgaa ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga
     4501 cgggcgttcc ttgcgcagct gtgctcgacg ttgtcactga agcgggaagg gactggctgc
     4561 tattgggcga agtgccgggg caggatctcc tgtcatctca ccttgctcct gccgagaaag
     4621 tatccatcat ggctgatgca atgcggcggc tgcatacgct tgatccggct acctgcccat
     4681 tcgaccacca agcgaaacat cgcatcgagc gagcacgtac tcggatggaa gccggtcttg
     4741 tcgatcagga tgatctggac gaagagcatc aggggctcgc gccagccgaa ctgttcgcca
     4801 ggctcaaggc gagcatgccc gacggcgagg atctcgtcgt gacccatggc gatgcctgct
     4861 tgccgaatat catggtggaa aatggccgct tttctggatt catcgactgt ggccggctgg
     4921 gtgtggcgga ccgctatcag gacatagcgt tggctacccg tgatattgct gaagagcttg
     4981 gcggcgaatg ggctgaccgc ttcctcgtgc tttacggtat cgccgctccc gattcgcagc
     5041 gcatcgcctt ctatcgcctt cttgacgagt tcttctgagc gggactctgg ggttcgaaat
     5101 gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta
     5161 tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg
     5221 ggatctcatg ctggagttct tcgcccaccc tagggggagg ctaactgaaa cacggaagga
     5281 gacaataccg gaaggaaccc gcgctatgac ggcaataaaa agacagaata aaacgcacgg
     5341 tgttgggtcg tttgttcata aacgcggggt tcggtcccag ggctggcact ctgtcgatac
     5401 cccaccgaga ccccattggg gccaatacgc ccgcgtttct tccttttccc caccccaccc
     5461 cccaagttcg ggtgaaggcc cagggctcgc agccaacgtc ggggcggcag gccctgccat
     5521 agcctcaggt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     5581 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     5641 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     5701 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     5761 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     5821 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     5881 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     5941 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     6001 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     6061 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     6121 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     6181 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     6241 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     6301 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     6361 tgtggataac cgtattaccg ccatgcat
//
