LOCUS       Exported                6064 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Hep C non-structural protein 5A (binds PI4KA).
ACCESSION   .
VERSION     .
KEYWORDS    NS5A-mCherry
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6064)
  AUTHORS   Hammond GR, Machner MP, Balla T
  TITLE     A novel probe for phosphatidylinositol 4-phosphate reveals multiple 
            pools beyond the Golgi.
  JOURNAL   J Cell Biol. 2014 Apr 7.
  PUBMED    24711504
REFERENCE   2  (bases 1 to 6064)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6064
                     /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      2014..2023
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             2020..2730
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELYK"
     primer_bind     complement(2165..2183)
                     /label=mCherry-R
                     /note="mCherry, reverse primer"
     primer_bind     2433..2452
                     /label=mCherry-F
                     /note="mCherry, forward primer"
     polyA_signal    2852..2973
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2889..2908)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2943..2962
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2980..3435)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3117..3138)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3329..3348
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3462..3566
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3568..3925
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3591..3611)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3776..3911
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3838..3857
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3960..4754
                     /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(4014..4033)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4624..4643
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4942..4961)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4986..5033
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5362..5950
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5851..5870
                     /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 ccggactcag atctcgagct caagcttacc atggcgtccg gctcgtggct aagggatgtt
      661 tgggattgga tatgcacggt gttgactgac ttcaagacct ggctccagtc caaactcctg
      721 ccgcggttac cgggagtccc tttcctgtca tgccaacgcg ggtacaaggg agtctggcgg
      781 ggggacggca tcatgcaaac cacctgccca tgcggagcac agatcgccgg acatgtcaaa
      841 aacggttcca tgaggatcgt agggcctaga acctgcagca acacgtggca cggaacgttc
      901 cccatcaacg catacaccac gggaccttgc acaccctccc cggcgcccaa ctattccagg
      961 gcgctatggc gggtggctgc tgaggagtac gtggaggtta cgcgtgtggg ggatttccac
     1021 tacgtgacgg gcatgaccac tgacaacgta aagtgcccat gccaggttcc ggcccccgaa
     1081 ttcttcacgg aggtggatgg agtgcggttg cacaggtacg ctccggcgtg caaacctctt
     1141 ctacgggagg acgtcacgtt ccaggtcggg ctcaaccaat acttggtcgg gtcgcagctc
     1201 ccatgcgagc ccgaaccgga cgtaacagtg cttacttcca tgctcaccga tccctcccac
     1261 attacagcag agacggctaa gcgtaggctg gctagagggt ctcccccctc tttagccagc
     1321 tcatcagcta gccagttgtc tgcgccttct ttgaaggcga catgcactac ccaccatgac
     1381 tccccggacg ctgacctcat cgaggccaac ctcttgtggc ggcaggagat gggcggaaac
     1441 atcactcgcg tggagtcaga gaataaggta gtaattctgg actctttcga accgcttcac
     1501 gcggaggggg atgagaggga gatatccgtc gcggcggaga tcctgcgaaa atccaggaag
     1561 ttcccctcag cgttgcccat atgggcacgc ccggactaca atcctccact gctagagtcc
     1621 tggaaggacc cggactacgt ccctccggtg gtacacggat gcccattgcc acctaccaag
     1681 gctcctccaa taccacctcc acggagaaag aggacggttg tcctgacaga atccaatgtg
     1741 tcttctgcct tggcggagct cgccactaag accttcggta gctccggatc gtcggccgtt
     1801 gatagcggca cggcgaccgc ccttcctgac ctggcctccg acgacggtga caaaggatcc
     1861 gacgttgagt cgtactcctc catgcccccc cttgaagggg agccggggga ccccgatctc
     1921 agcgacgggt cttggtctac cgtgagtgag gaggctagtg aggatgtcgt ctgctgccag
     1981 tcgacggtac cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag
     2041 gataacatgg ccatcatcaa ggagttcatg cgcttcaagg tgcacatgga gggctccgtg
     2101 aacggccacg agttcgagat cgagggcgag ggcgagggcc gcccctacga gggcacccag
     2161 accgccaagc tgaaggtgac caagggtggc cccctgccct tcgcctggga catcctgtcc
     2221 cctcagttca tgtacggctc caaggcctac gtgaagcacc ccgccgacat ccccgactac
     2281 ttgaagctgt ccttccccga gggcttcaag tgggagcgcg tgatgaactt cgaggacggc
     2341 ggcgtggtga ccgtgaccca ggactcctcc ctgcaggacg gcgagttcat ctacaaggtg
     2401 aagctgcgcg gcaccaactt cccctccgac ggccccgtaa tgcagaagaa gaccatgggc
     2461 tgggaggcct cctccgagcg gatgtacccc gaggacggcg ccctgaaggg cgagatcaag
     2521 cagaggctga agctgaagga cggcggccac tacgacgctg aggtcaagac cacctacaag
     2581 gccaagaagc ccgtgcagct gcccggcgcc tacaacgtca acatcaagtt ggacatcacc
     2641 tcccacaacg aggactacac catcgtggaa cagtacgaac gcgccgaggg ccgccactcc
     2701 accggcggca tggacgagct gtacaagtaa gcggccgcga ctctagatca taatcagcca
     2761 taccacattt gtagaggttt tacttgcttt aaaaaacctc ccacacctcc ccctgaacct
     2821 gaaacataaa atgaatgcaa ttgttgttgt taacttgttt attgcagctt ataatggtta
     2881 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     2941 ttgtggtttg tccaaactca tcaatgtatc ttaaggcgta aattgtaagc gttaatattt
     3001 tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa
     3061 tcggcaaaat cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag
     3121 tttggaacaa gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg
     3181 tctatcaggg cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga
     3241 ggtgccgtaa agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg
     3301 gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg
     3361 cgctggcaag tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc
     3421 cgctacaggg cgcgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     3481 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     3541 ttcaataata ttgaaaaagg aagagtcctg aggcggaaag aaccagctgt ggaatgtgtg
     3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3841 ttatgcagag gccgaggccg cctcggcctc tgagctattc cagaagtagt gaggaggctt
     3901 ttttggaggc ctaggctttt gcaaagatcg atcaagagac aggatgagga tcgtttcgca
     3961 tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag aggctattcg
     4021 gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc cggctgtcag
     4081 cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg aatgaactgc
     4141 aagacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc gcagctgtgc
     4201 tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg ccggggcagg
     4261 atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct gatgcaatgc
     4321 ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg aaacatcgca
     4381 tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat ctggacgaag
     4441 agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgagc atgcccgacg
     4501 gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg gtggaaaatg
     4561 gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc tatcaggaca
     4621 tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct gaccgcttcc
     4681 tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat cgccttcttg
     4741 acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga cgcccaacct
     4801 gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct tcggaatcgt
     4861 tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg agttcttcgc
     4921 ccaccctagg gggaggctaa ctgaaacacg gaaggagaca ataccggaag gaacccgcgc
     4981 tatgacggca ataaaaagac agaataaaac gcacggtgtt gggtcgtttg ttcataaacg
     5041 cggggttcgg tcccagggct ggcactctgt cgatacccca ccgagacccc attggggcca
     5101 atacgcccgc gtttcttcct tttccccacc ccacccccca agttcgggtg aaggcccagg
     5161 gctcgcagcc aacgtcgggg cggcaggccc tgccatagcc tcaggttact catatatact
     5221 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     5281 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     5341 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     5401 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     5461 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     5521 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     5581 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     5641 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     5701 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     5761 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     5821 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     5881 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     5941 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     6001 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgccat
     6061 gcat
//
