LOCUS       40924_41165        8867 bp DNA     circular SYN 18-DEC-2018
DEFINITION  Cloning vector pSplPIGver2, complete sequence.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8867)
  AUTHORS   Gurskaia NG, Staroverov DB, Fradkov AF, Luk'ianov KA.
  TITLE     [Coding region of far-red fluorescent protein katushka contains a 
            strong donor splice site]
  JOURNAL   Bioorg. Khim. 37 (3), 425-428 (2011)
  PUBMED    21899059
REFERENCE   2  (bases 1 to 8867)
  AUTHORS   Gurskaya NG, Staroverov DB, Zhang L, Fradkov AF, Markina NM, 
            Pereverzev AP, Lukyanov KA.
  TITLE     Analysis of alternative splicing of cassette exons at single-cell 
            level using two fluorescent proteins
  JOURNAL   Nucleic Acids Res. 40 (8), E57 (2012)
  PUBMED    22259036
REFERENCE   3  (bases 1 to 8867)
  AUTHORS   Gurskaya NG, Staroverov DB, Fradkov AF, Markina NM, Perverzev AP, 
            Lukyanov KA.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-SEP-2015) Department of Genetics and Postgenomic 
            Technologies Laboratory of Biophotonics, M.M. Shemyakin and Yu.a. 
            Ovchinnikov Institute of Bioorganic Chemistry of the Russian Academy
            of Sciences, Ulitsa Miklukho-Maklaya, 16/10, Moscow 117997, Russian 
            Federation
REFERENCE   4  (bases 1 to 8867)
  TITLE     Direct Submission
REFERENCE   5  (bases 1 to 8867)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Bioorg. 
            Khim."; date: "2011"; volume: "37"; issue: "3"; pages: "425-428"
COMMENT     SGRef: number: 2; type: "Journal Article"; journalName: "Nucleic 
            Acids Res."; date: "2012"; volume: "40"; issue: "8"; pages: "E57"
COMMENT     SGRef: number: 3; type: "Journal Article"; journalName: "Submitted 
            (28-SEP-2015) Department of Genetics and Postgenomic Technologies 
            Laboratory of Biophotonics, M.M. Shemyakin and Yu.a. Ovchinnikov 
            Institute of Bioorganic Chemistry of the Russian Academy of 
            Sciences, Ulitsa Miklukho-Maklaya, 16/10, Moscow 117997, Russian 
            Federation"
COMMENT     SGRef: number: 4; type: "Journal Article"
COMMENT     ##Assembly-Data-START##
            Sequencing Technology :: Sanger dideoxy sequencing 
            ##Assembly-Data-END##
FEATURES             Location/Qualifiers
     source          1..8867
                     /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"
     misc_feature    590..612
                     /note="multiple cloning site; enhancer; TATA box"
     CDS             613..1317
                     /label=TurboFP635
                     /note="far-red-shifted derivative of red fluorescent
                     protein from Entacmaea quadricolor, also known as Katushka 
                     (Shcherbo et al., 2007)"
     misc_feature    1318..1333
                     /label=multiple cloning site
                     /note="multiple cloning site"
     misc_feature    4790..4816
                     /label=multiple cloning site
                     /note="multiple cloning site"
     CDS             4817..5530
                     /label=TagGFP2
                     /note="monomeric green fluorescent protein, also known as 
                     mTagGFP"
     polyA_signal    5655..5776
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(5783..6238)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6265..6369
                     /label=AmpR promoter
     promoter        6371..6728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             6763..7554
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
     polyA_signal    7789..7836
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      8165..8753
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
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 ccatggtggg tgaggatagc gtgctgatca ccgagaacat gcacatgaaa
      661 ctgtacatgg agggcaccgt gaacgaccac cacttcaagt gcacatccga gggcgaaggc
      721 aagccctacg agggcaccca gaccatgaag atcaaggtgg tcgagggcgg ccctctcccc
      781 ttcgccttcg acatcctggc taccagcttc atgtacggca gcaaaacctt tatcaaccac
      841 acccagggca tccccgactt ctttaagcag tccttccctg agggcttcac atgggagagg
      901 atcaccacat acgaagacgg gggcgtgctg accgctaccc aggacaccag cctccagaac
      961 ggctgcctca tctacaacgt caagatcaac ggggtgaact tcccatccaa cggccctgtg
     1021 atgcagaaga aaacactcgg ctgggaggcc agcaccgaga tgctgtaccc cgctgacagc
     1081 ggcctgagag gccatagcca gatggccctg aagctcgtgg gcgggggcta cctgcactgc
     1141 tccctcaaga ccacatacag atccaagaaa cccgctaaga acctcaagat gcccggcttc
     1201 tacttcgtgg acaggagact ggaaagaatc aaggaggccg acaaagagac ctacgtcgag
     1261 cagcacgaga tggctgtggc tcgctactgc gacctgccta gcaaactggg gcacagctcc
     1321 ggactcaaga tctccggaaa tgttcaggct ggagactatg tgctaatcca tgcaggactg
     1381 agtggtgtgg gcacagctgc tatccaactc acccggatgg ctggagctat tcctctggtc
     1441 acagctggct cccagaagaa gcttcaaatg gcagaaaagc ttggagcagc tgctggattc
     1501 aattacaaaa aagaggattt ctctgaagca acgctgaaat tcaccaaagg taaacaactg
     1561 cttctacagt tcagcaggaa tttaagagat gattaaataa aagccccacc agtctcaaag
     1621 ttgccagtgc tgtgtatcag tcttacctct gctaagtgca caacagctgc cctggccaaa
     1681 ttctagtact atggtatttg accactggag acaggcagtc atatttaaat ctaagatggt
     1741 ttccatgtgc attcttgcaa gagataagat tcatccacac tttagaccat gcagacttgg
     1801 tctggaatta tccagcacct tttcttccca accagccatt ccacacttag ttaacaggag
     1861 gcttggaggc agagaccagt ttggagccag ccatggtagg gttgggtggt agcaccctaa
     1921 atgagtaagt agatgtgcac acctctgtat tccaaagcca gacgtgtcta tttttggctc
     1981 atccctttcc gtttctcttt cttttttttt tttgagatgg agtctcactc tgtcactgag
     2041 gatggcgtgc agtggcttga tctcggctcg ctgcaacctc cgcctcccag gttcaagtga
     2101 ttctccagcc ttagcctcct gagtagctga gatcacaggc gcaccaccac gcccagctaa
     2161 tttttgtatt tttagtagag acgggctttc actatgttgg ccaggctggt ctcgaactcc
     2221 tgacctcagg tgatccacct gcctcagcct cccaaagtac tgggattaca ggcatgagcc
     2281 accatgcctg gcctccattt tttatgtgag tagaaaagga gagatgaaac caaagctaga
     2341 aagcctatga ttccaataac ctagtgcgga cacttcctaa cttctatgtg aacaactagc
     2401 ctaggttggg cctcctgtat tctgttctca atcgtgcctg ggtgtcagtg ggactgtact
     2461 tattttggca tgacaggaga cgctaggtca tggtgcccca tttccctctt gctgtagcta
     2521 ggcctcataa atggtgaact tcatacatgc tatcatagag ttctgctatc taaactccat
     2581 ttacaacagt tcttcaagca gtgtttctaa ttccaaccta tagatttacc acttgcatct
     2641 gtatttatat ccataatccc caaatatacc atggtatctt atttagcacc aagctgacta
     2701 gcatcacaaa tctgtttcct atggaaggtg ctggagttaa tcttattcta gactgcatag
     2761 gcggatccta ctgggagaag aacgtcaact gcctggctct tgatggtcga tgggttctct
     2821 atggtctgat gggaggaggt gacatcaatg ggcccctgtt ttcaaagcta ctttttaagc
     2881 gaggaagtct gatcaccagt ttgctgaggt ctagggacaa taaggtgagt gatgaatgaa
     2941 aagaaaacat gtgatttaac ccagaaaagt gtggttcatt tacattaccc cgagaaagga
     3001 agctctacct tagattacgg aggttagttt ctatgaagaa acctccccct ctgaagataa
     3061 agagggaacc cgatttcaga gaactgattc agattggttg atagaagctt cagtatcatg
     3121 aaaagaagtg tagctgatac ttcacaggtt atctgaacat tctaacagaa tgaagaaagc
     3181 actgactata ttaggaagta tttcgagtag ttccaagact ttgttttcaa aggcagaatt
     3241 catctcagtg tgctagaaag tctcttaggt gagtaaaaaa ataagtaagc cttggccttc
     3301 aacagattat aggttccatg ccgggcacag tggcacttgc ttgtagtccc agctacttgg
     3361 aaggctgagg cagaggattg cttgagtcca ggagtttcaa ggctgcagtg agctatgatt
     3421 gcacctgtgc atagccactg taccccagcc tgagcaacat agcaaaacct catctcaaaa
     3481 aaaaaaagat tatatgctcc tgaaatattc catctggcag tctagcaggt accacctttg
     3541 ggaatgtttc caccataggg ttttgcacca gctgagggac aggagagctt aagcatcctt
     3601 ttttcactaa gcctatatga gtcctatctg tggctgcctc ctgggtaaag acactttatt
     3661 taaaaacaat tcttaggctg tcagtgacac taacatttta ctccaggctt cagtctttat
     3721 ctggtacaga acagaagtta cacatttgga ggtacaaggg agccagagat cacctgtaac
     3781 ccccctttag ctcctatgtg gtctcttcag ctggctctag aaaccaaatt gataccagca
     3841 gattgacagg agaaaagcat gtactttttt tttttttttt tttttttatg gagacagggt
     3901 cttactctgt cacccaggct ggagtgcaat ggcacaatca gcctcctggg tagctaagac
     3961 tgaaggcata ccaccatgcc tggcttgtat tttttataga gacaggtttt caccatgttg
     4021 cccaggctga tctcaaactc ccaggctcaa gtgattcggc caaagtaaga tgcttttgta
     4081 atttttagac aaagaacaat aaatttgaga agaaataaga caaaatctgg ctagggatag
     4141 taaatttcta gggaagtcac taggagatat atgggggtgg ggggtgtaaa actagtggaa
     4201 gataagggtt acttccttaa gtgtatttat tcagttccat tgcagccccg tttccagtct
     4261 ctggtgataa gggctttttg ccctagtatg gggagggtac ccctctcaga ggaatcttta
     4321 tgacttgcta catgcaagaa gagacaggtc agctagcctt ttctgaaact acaatttctc
     4381 caatgctttc acctcaaaat taacaatata cctatcggac atattctgag atggcacatg
     4441 actccttcaa gggtatttca cacacatgct aggaagtgag gggttaagtg tccctgtgtg
     4501 ctttttcaga tacgtggctt cagagagagc aagaaggctg aggaggatga ggagggcagg
     4561 gtgaggctct ctccccaggc tgatgatctc ctttcccttg tcttagtaca agcaaatgct
     4621 ggtgaatgct ttcacggagc aaattctgcc tcacttctcc acggagggcc cccaacgtct
     4681 gctgccggtt ctggacagaa tctacccagt gaccgaaatc caggaggccc ataagtacat
     4741 ggaggccaac aagaacatag gcaagatcgt cctggaactg ccccagacgg tcgaccggga
     4801 tccaccggtc gccaccatga gcgggggcga ggagctgttc gccggcatcg tgcccgtgct
     4861 gatcgagctg gacggcgacg tgcacggcca caagttcagc gtgcgcggcg agggcgaggg
     4921 cgacgccgac tacggcaagc tggagatcaa gttcatctgc accaccggca agctgcccgt
     4981 gccctggccc accctggtga ccaccctctg ctacggcatc cagtgcttcg cccgctaccc
     5041 cgagcacatg aagatgaacg acttcttcaa gagcgccatg cccgagggct acatccagga
     5101 gcgcaccatc cagttccagg acgacggcaa gtacaagacc cgcggcgagg tgaagttcga
     5161 gggcgacacc ctggtgaacc gcatcgagct gaagggcaag gacttcaagg aggacggcaa
     5221 catcctgggc cacaagctgg agtacagctt caacagccac aacgtgtaca tccgccccga
     5281 caaggccaac aacggcctgg aggctaactt caagacccgc cacaacatcg agggcggcgg
     5341 cgtgcagctg gccgaccact accagaccaa cgtgcccctg ggcgacggcc ccgtgctgat
     5401 ccccatcaac cactacctga gcactcagac caagatcagc aaggaccgca acgaggcccg
     5461 cgaccacatg gtgctcctgg agtccttcag cgcctgctgc cacacccacg gcatggacga
     5521 gctgtacagg taagcggccg cgactctaga tcataatcag ccataccaca tttgtagagg
     5581 ttttacttgc tttaaaaaac ctcccacacc tccccctgaa cctgaaacat aaaatgaatg
     5641 caattgttgt tgttaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     5701 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     5761 tcatcaatgt atcttaacgc gtaaattgta agcgttaata ttttgttaaa attcgcgtta
     5821 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
     5881 aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa caagagtcca
     5941 ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc
     6001 ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg taaagcacta
     6061 aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc ggcgaacgtg
     6121 gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg
     6181 gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca gggcgcgtca
     6241 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     6301 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     6361 aggaagagtc ctgaggcgga aagaaccagc tgtggaatgt gtgtcagtta gggtgtggaa
     6421 agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     6481 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     6541 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     6601 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     6661 ccgcctcggc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     6721 tttgcaaaga tcgatcaaga gacaggatga ggatcgtttc gcatgattga acaagatgga
     6781 ttgcacgcag gttctccggc cgcttgggtg gagaggctat tcggctatga ctgggcacaa
     6841 cagacaatcg gctgctctga tgccgccgtg ttccggctgt cagcgcaggg gcgcccggtt
     6901 ctttttgtca agaccgacct gtccggtgcc ctgaatgaac tgcaagacga ggcagcgcgg
     6961 ctatcgtggc tggccacgac gggcgttcct tgcgcagctg tgctcgacgt tgtcactgaa
     7021 gcgggaaggg actggctgct attgggcgaa gtgccggggc aggatctcct gtcatctcac
     7081 cttgctcctg ccgagaaagt atccatcatg gctgatgcaa tgcggcggct gcatacgctt
     7141 gatccggcta cctgcccatt cgaccaccaa gcgaaacatc gcatcgagcg agcacgtact
     7201 cggatggaag ccggtcttgt cgatcaggat gatctggacg aagagcatca ggggctcgcg
     7261 ccagccgaac tgttcgccag gctcaaggcg agcatgcccg acggcgagga tctcgtcgtg
     7321 acccatggcg atgcctgctt gccgaatatc atggtggaaa atggccgctt ttctggattc
     7381 atcgactgtg gccggctggg tgtggcggac cgctatcagg acatagcgtt ggctacccgt
     7441 gatattgctg aagagcttgg cggcgaatgg gctgaccgct tcctcgtgct ttacggtatc
     7501 gccgctcccg attcgcagcg catcgccttc tatcgccttc ttgacgagtt cttctgagcg
     7561 ggactctggg gttcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     7621 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     7681 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccaccct agggggaggc
     7741 taactgaaac acggaaggag acaataccgg aaggaacccg cgctatgacg gcaataaaaa
     7801 gacagaataa aacgcacggt gttgggtcgt ttgttcataa acgcggggtt cggtcccagg
     7861 gctggcactc tgtcgatacc ccaccgagac cccattgggg ccaatacgcc cgcgtttctt
     7921 ccttttcccc accccacccc ccaagttcgg gtgaaggccc agggctcgca gccaacgtcg
     7981 gggcggcagg ccctgccata gcctcaggtt actcatatat actttagatt gatttaaaac
     8041 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     8101 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     8161 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     8221 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     8281 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     8341 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     8401 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     8461 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     8521 cgacctacac cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg
     8581 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     8641 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     8701 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     8761 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     8821 ctgcgttatc ccctgattct gtggataacc gtattaccgc catgcat
//
