LOCUS       Exported                5586 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Localization: Membrane Scaffold, Excitation: 400 / 490, Emission: 
            468 / 511.
ACCESSION   .
VERSION     .
KEYWORDS    PS-CFP2-PCNA-19-SV40NLS-4
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5586)
  TITLE     Davidson Photoactivatable Fluorescent Proteins
REFERENCE   2  (bases 1 to 5586)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5586
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     enhancer        1..304
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        305..508
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     459..479
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      535..544
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             547..567
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             571..591
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             595..615
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             628..1341
                     /codon_start=1
                     /product="photoswitchable cyan-to-green fluorescent 
                     protein"
                     /label=PS-CFP2
                     /note="mammalian codon-optimized"
                     /translation="MSKGAELFTGIVPILIELNGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVATLSYGVQCFSRYPDHMKQHDFFKSAMPEGYIQERTIFFEDDG
                     NYKTRAEVKFEGDTLVSRIELTGTDFKEDGNILGNKMEYNYNATNVYIVADKARNGIKV
                     NFKVRHNIKDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMIYLE
                     FVTAAAITHGMDELYK"
     polyA_signal    2314..2435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2351..2370)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2405..2424
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2442..2897)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2579..2600)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2791..2810
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        2924..3028
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3030..3387
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3053..3073)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3238..3373
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3300..3319
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3422..4216
                     /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(3476..3495)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4086..4105
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4404..4423)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4448..4495
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      4824..5412
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5313..5332
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
       61 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      121 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      181 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      241 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      301 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      361 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      421 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      481 acggtgggag gtctatataa gcagagctgg tttagtgaac cgtcagatcc gctagccacc
      541 atggatccaa aaaagaagag aaaggtagat ccaaaaaaga agagaaaggt agatccaaaa
      601 aagaagagaa aggtaccggt cgccaccatg agcaagggcg ccgagctgtt caccggcatc
      661 gtgcccatcc tgatcgagct gaatggcgat gtgaatggcc acaagttcag cgtgagcggc
      721 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
      781 aagctgcctg tgccctggcc caccctggtg gccaccctga gctacggcgt gcagtgcttc
      841 tcacgctacc ccgatcacat gaagcagcac gacttcttca agagcgccat gcctgagggc
      901 tacatccagg agcgcaccat cttcttcgag gatgacggca actacaagac gcgcgccgag
      961 gtgaagttcg agggcgatac cctggtgagt cgcatcgagc tgaccggcac tgatttcaag
     1021 gaggatggca acatcctggg caataagatg gagtacaact acaacgccac caatgtgtac
     1081 atcgtggccg acaaggccag gaatggcatc aaggtgaact tcaaggtccg ccacaacatc
     1141 aaggatggca gcgtgcagct ggccgaccac taccagcaga atacccccat cggcgatggc
     1201 cctgtgctgc tgcccgataa ccactacctg tccacccaga gcgccctgtc caaggacccc
     1261 aacgagaagc gcgatcacat gatctacctc gagttcgtga ccgccgccgc catcacccac
     1321 ggcatggatg agctgtacaa gtccggagaa gggcaagggc aagggcaagg gccgggccgc
     1381 ggctacgcgt atcgatccat gttcgaggcg cgcctggtcc agggctccat cctcaagaag
     1441 gtgttggagg cactcaagga cctcatcaac gaggcctgct gggatattag ctccagcggt
     1501 gtaaacctgc agagcatgga ctcgtcccac gtctctttgg tgcagctcac cctgcggtct
     1561 gagggcttcg acacctaccg ctgcgaccgc aacctggcca tgggcgtgaa cctcaccagt
     1621 atgtccaaaa tactaaaatg cgccggcaat gaagatatca ttacactaag ggccgaagat
     1681 aacgcggata ccttggcgct agtatttgaa gcaccaaacc aggagaaagt ttcagactat
     1741 gaaatgaagt tgatggattt agatgttgaa caacttggaa ttccagaaca ggagtacagc
     1801 tgtgtagtaa agatgccttc tggtgaattt gcacgtatat gccgagatct cagccatatt
     1861 ggagatgctg ttgtaatttc ctgtgcaaaa gacggagtga aattttctgc aagtggagaa
     1921 cttggaaatg gaaacattaa attgtcacag acaagtaatg tcgataaaga ggaggaagct
     1981 gttaccatag agatgaatga accagttcaa ctaacttttg cactgaggta cctgaacttc
     2041 tttacaaaag ccactccact ctcttcaacg gtgacactca gtatgtctgc agatgtaccc
     2101 cttgttgtag agtataaaat tgcggatatg ggacacttaa aatactactt ggctcccaag
     2161 atcgaggatg aagaaggatc ttagggatcc accggatcta gataactgat cataatcagc
     2221 cataccacat ttgtagaggt tttacttgct ttaaaaaacc tcccacacct ccccctgaac
     2281 ctgaaacata aaatgaatgc aattgttgtt gttaacttgt ttattgcagc ttataatggt
     2341 tacaaataaa gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct
     2401 agttgtggtt tgtccaaact catcaatgta tcttaacgcg taaattgtaa gcgttaatat
     2461 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     2521 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     2581 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     2641 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     2701 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     2761 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     2821 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     2881 gccgctacag ggcgcgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     2941 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3001 gcttcaataa tattgaaaaa ggaagagtcc tgaggcggaa agaaccagct gtggaatgtg
     3061 tgtcagttag ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg
     3121 catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt
     3181 atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc
     3241 ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt
     3301 atttatgcag aggccgaggc cgcctcggcc tctgagctat tccagaagta gtgaggaggc
     3361 ttttttggag gcctaggctt ttgcaaagat cgatcaagag acaggatgag gatcgtttcg
     3421 catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt
     3481 cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     3541 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     3601 gcaagacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     3661 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     3721 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     3781 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     3841 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     3901 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcga gcatgcccga
     3961 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     4021 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     4081 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     4141 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     4201 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     4261 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     4321 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     4381 gcccacccta gggggaggct aactgaaaca cggaaggaga caataccgga aggaacccgc
     4441 gctatgacgg caataaaaag acagaataaa acgcacggtg ttgggtcgtt tgttcataaa
     4501 cgcggggttc ggtcccaggg ctggcactct gtcgataccc caccgagacc ccattggggc
     4561 caatacgccc gcgtttcttc cttttcccca ccccaccccc caagttcggg tgaaggccca
     4621 gggctcgcag ccaacgtcgg ggcggcaggc cctgccatag cctcaggtta ctcatatata
     4681 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     4741 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     4801 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     4861 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     4921 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg
     4981 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5041 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     5101 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     5161 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga
     5221 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     5281 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     5341 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     5401 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     5461 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     5521 atgcattagt tattaatagt aatcaattac ggggtcatta gttcatagcc catatatgga
     5581 gttccg
//
