LOCUS       Exported                6404 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Expression vector Influenza A virus FPV (H7N1) hemagglutinin mEGFP 
            fusion protein, plasma membrane localization.
ACCESSION   .
VERSION     .
KEYWORDS    Influenza-FPV-HA-mEGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6404)
  AUTHORS   Dunsing V, Luckner M, Zuhlke B, Petazzi RA, Herrmann A, Chiantia S
  TITLE     Optimal fluorescent protein tags for quantifying protein 
            oligomerization in living cells.
  JOURNAL   Sci Rep. 2018 Jul 13;8(1):10634. doi: 10.1038/s41598-018-28858-0.
  PUBMED    30006597
REFERENCE   2  (bases 1 to 6404)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6404
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        1..204
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     155..175
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     regulatory      1980..1989
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             1986..2705
                     /codon_start=1
                     /product="enhanced GFP with monomerizing A206K mutation 
                     (Zacharias et al., 2002)"
                     /label=mEGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSKLSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2031..2052)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2292..2311)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2639..2660
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2828..2949
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2865..2884)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2919..2938
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     rep_origin      complement(2956..3411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3093..3114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3305..3324
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        3438..3542
                     /gene="bla"
                     /label=AmpR promoter
     promoter        3544..3901
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     primer_bind     complement(3567..3587)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      3752..3887
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3814..3833
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             3936..4730
                     /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(3990..4009)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4600..4619
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     complement(4918..4937)
                     /label=TK-pA-R
                     /note="Thymidine kinase polyA, reverse primer"
     polyA_signal    4962..5009
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5338..5926
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5827..5846
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     enhancer        6101..6404
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtgatgcggt tttggcagta catcaatggg cgtggatagc ggtttgactc acggggattt
       61 ccaagtctcc accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac
      121 tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg
      181 tgggaggtct atataagcag agctggttta gtgaaccgtc agatccgcta gcgctaccgg
      241 actcagatct catgaacact caaatcctgg ttttcgccct tgtggcagtc attcccacaa
      301 atgcagacaa aatttgtctt ggacatcatg ctgtatcaaa tggcaccaaa gtaaacacac
      361 tcactgagag aggagtagaa gttgtcaatg caacggaaac agtggagcgg acaaacatcc
      421 ccaaaatttg ctcaaaaggg aaaagaacca ctgatcttgg ccaatgcgga ctgttaggga
      481 ccattaccgg accacctcaa tgcgaccaat ttctagaatt ttcagctgat ctaataatcg
      541 agagacgaga aggaaatgat gtttgttacc cgggaaagtt tgttaatgaa gaggcattgc
      601 ggcaaatcct cagaggatca ggtgggattg acaaagaaac aatgggattc acatatagtg
      661 gaataaggac caacggaaca actagtgcat gtagaagatc agggtcttca ttctatgcag
      721 aaatggagtg gctcctgtca aatacagaca atgcttcttt cccacaaatg acaaaatcat
      781 acaaaaacac aaggagagaa tcagctctga tagtatgggg aatccaccat tcaggatcaa
      841 ccaccgaaca gaccaaacta tatgggagtg gaaataaact gataacagtc gggagttcca
      901 aatatcatca atcttttgtg ccgagtccag gaacacgacc gcagataaat ggccagtccg
      961 gacggattga ttttcattgg ttgatcttgg atcccaatga tacagttact tttagtttca
     1021 atggggcttt catagctcca aatcgtgcca gcttcttgag gggaaagtcc atggggatcc
     1081 agagcgatgt gcaggttgat gccaattgcg aaggggaatg ctaccacagt ggagggacta
     1141 taacaagcag attgcctttt caaaacatca atagcagagc agttggcaaa tgcccaagat
     1201 atgtaaaaca ggaaagttta ttattggcaa ctgggatgaa gaacgttccc gaaccttcca
     1261 aaaaaggcaa aaaaagaggc ctgtttggcg ctatagcagg gtttattgaa aatggttggg
     1321 aaggtctggt cgacgggtgg tacggtttca ggcatcagaa tgcacaagga gaaggaactg
     1381 cagcagacta caaaagcacc caatcggcaa ttgatcagat aaccggaaag ttaaatagac
     1441 tcattgagaa aaccaaccag caatttgagc taatagataa tgaattcact gaggtggaaa
     1501 agcagattgg caatttaatt aactggacca aagactccat cacagaagta tggtcttaca
     1561 atgctgaact tattgtggca atggaaaacc agcacactat tgatttggcc gattcagaga
     1621 tgaacaggct gtatgagcga gtgaggaaac aattaaggga aaatgctgaa gaggatggca
     1681 ctggttgctt tgaaattttt cataaatgtg acgatgattg tatggctagt ataaggaaca
     1741 atacttatga tcacagcaaa tacagagaag aagcgatgca aaatagaata caaattgacc
     1801 cagtcaaatt gagtagtggc tacaaagatg tgatactttg gtttagcttc ggggcatcat
     1861 gctttttgct tcttgccatt gcaatgggcc ttgttttcat atgtgtgaag aacggaaaca
     1921 tgcggtgcac tatttgtata ctccggcctg aagctccgcg ggcccgggat ccaccggtcg
     1981 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc
     2041 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca
     2101 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc
     2161 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca
     2221 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca
     2281 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca
     2341 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg
     2401 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga
     2461 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc
     2521 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca
     2581 accactacct gagcacccag tccaagctga gcaaagaccc caacgagaag cgcgatcaca
     2641 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca
     2701 agtaaagcgg ccgcgactct agatcataat cagccatacc acatttgtag aggttttact
     2761 tgctttaaaa aacctcccac acctccccct gaacctgaaa cataaaatga atgcaattgt
     2821 tgttgttaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2881 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2941 tgtatcttaa ggcgtaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt
     3001 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
     3061 aagaatagac cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa
     3121 agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac
     3181 gtgaaccatc accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga
     3241 accctaaagg gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa
     3301 aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc
     3361 tgcgcgtaac caccacaccc gccgcgctta atgcgccgct acagggcgcg tcaggtggca
     3421 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     3481 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     3541 gtcctgaggc ggaaagaacc agctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     3601 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     3661 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     3721 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     3781 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     3841 ggcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     3901 agatcgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg
     3961 caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa
     4021 tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg
     4081 tcaagaccga cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt
     4141 ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa
     4201 gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc
     4261 ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg
     4321 ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg
     4381 aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg
     4441 aactgttcgc caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg
     4501 gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga ttcatcgact
     4561 gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc cgtgatattg
     4621 ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt atcgccgctc
     4681 ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga gcgggactct
     4741 ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt tcgattccac
     4801 cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg gctggatgat
     4861 cctccagcgc ggggatctca tgctggagtt cttcgcccac cctaggggga ggctaactga
     4921 aacacggaag gagacaatac cggaaggaac ccgcgctatg acggcaataa aaagacagaa
     4981 taaaacgcac ggtgttgggt cgtttgttca taaacgcggg gttcggtccc agggctggca
     5041 ctctgtcgat accccaccga gaccccattg gggccaatac gcccgcgttt cttccttttc
     5101 cccaccccac cccccaagtt cgggtgaagg cccagggctc gcagccaacg tcggggcggc
     5161 aggccctgcc atagcctcag gttactcata tatactttag attgatttaa aacttcattt
     5221 ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta
     5281 acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg
     5341 agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc
     5401 ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag
     5461 cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa
     5521 gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc
     5581 cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc
     5641 gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta
     5701 caccgaactg agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag
     5761 aaaggcggac aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct
     5821 tccaggggga aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga
     5881 gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc
     5941 ggccttttta cggttcctgg ccttttgctg gccttttgct cacatgttct ttcctgcgtt
     6001 atcccctgat tctgtggata accgtattac cgccatgcat tagttattaa tagtaatcaa
     6061 ttacggggtc attagttcat agcccatata tggagttccg cgttacataa cttacggtaa
     6121 atggcccgcc tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg
     6181 ttcccatagt aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt
     6241 aaactgccca cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg
     6301 tcaatgacgg taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc
     6361 ctacttggca gtacatctac gtattagtca tcgctattac catg
//
