LOCUS       Exported                7915 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgA1) high 
            affinity variant 2.
ACCESSION   .
VERSION     .
KEYWORDS    pcDNA3-sACE2v2-Fc(IgA1)
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7915)
  AUTHORS   Chan KK, Dorosky D, Sharma P, Abbasi SA, Dye JM, Kranz DM, Herbert 
            AS, Procko E
  TITLE     Engineering human ACE2 to optimize binding to the spike protein of 
            SARS coronavirus 2.
  JOURNAL   Science. 2020 Sep 4;369(6508):1261-1265. doi: 
            10.1126/science.abc0870. Epub 2020 Aug 4.
  PUBMED    32753553
REFERENCE   2  (bases 1 to 7915)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7915
                     /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"
     primer_bind     249..268
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        249..267
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(2892..2909)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2898..3122
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3168..3596
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3255..3274)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3465..3486
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3605..3625)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3610..3939
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3790..3925
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3852..3871
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4006..4800
                     /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
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(4060..4079)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     4670..4689
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    4974..5095
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5011..5030)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5065..5084
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5144..5160)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5144..5160)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5157..5179)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5168..5184
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
     promoter        complement(5192..5222)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5237..5258
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5375..5392)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5546..6134)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5626..5645)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(6305..7165)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6928..6947
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7166..7270)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7345..7364)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        7536..7915
                     /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 agctctctgg ctaactagag aacccactgc ttactggctt
      241 atcgaaatta atacgactca ctatagggag acccaagctg gctagcgcca ccatgtcaag
      301 ctcttcctgg ctccttctca gccttgttgc tgtaactgct gctcagtcca ccattgagga
      361 acaggccaag tactttttgg acaagtttaa ccacgaagcc gaagacctgt tctatcaaag
      421 ttcacttgct tcttggaatt ataacaccaa tattactgaa gagaatgtcc aaaacatgaa
      481 taatgctggg gacaaatggt ctgccttttt aaaggaacag tccacaactg cccaaatgta
      541 tccactacaa gaaattcaga atctcacagt caagcttcag ctgcaggctc ttcagcaaaa
      601 tgggtcttca gtgctctcag aagacaagag caaacggttg aacacaattc taaatacaat
      661 gagcaccatc tacagtactg gaaaagtttg taacccagat aatccacaag aatgcttatt
      721 acttgaacca ggtttgaatg aaataatggc aaacagttta gactacaatg agaggctctg
      781 ggcttgggaa agctggagat ctgaggtcgg caagcagctg aggccattat atgaagagta
      841 tgtggtcttg aaaaatgaga tggcaagagc aaatcattat gaggactatg gggattattg
      901 gagaggagac tatgaagtaa atggggtaga tggctatgac tacagccgcg gccagttgat
      961 tgaagatgtg gaacatacct ttgaagagat taaaccatta tatgaacatc ttcatgccta
     1021 tgtgagggca aagttgatga atgcctatcc ttcctatatc agtccaattg gatgcctccc
     1081 tgctcatttg cttggtgata tgtggggtag attttggaca aatctgtact ctttgacagt
     1141 tccctttgga cagaaaccaa acatagatgt tactgatgca atggtggacc aggcctggga
     1201 tgcacagaga atattcaagg aggccgagaa gttctttgta tctgttggtc ttcctaatat
     1261 gactcaagga ttctgggaat attccatgct aacggaccca ggaaatgttc agaaagcagt
     1321 ctgccatccc acagcttggg acctggggaa gggcgacttc aggattctta tgtgcacaaa
     1381 ggtgacaatg gacgacttcc tgacagctca tcatgagatg gggcatatcc agtatgatat
     1441 ggcatatctt gcacaacctt ttctgctaag aaatggagct aatgaaggat tccatgaagc
     1501 tgttggggaa atcatgtcac tttctgcagc cacacctaag catttaaaat ccattggtct
     1561 tctgtcaccc gattttcaag aagacaatga aacagaaata aacttcctgc tcaaacaagc
     1621 actcacgatt gttgggactc tgccatttac ttacatgtta gagaagtgga ggtggatggt
     1681 ctttaaaggg gaaattccca aagaccagtg gatgaaaaag tggtgggaga tgaagcgaga
     1741 gatagttggg gtggtggaac ctgtgcccca tgatgaaaca tactgtgacc ccgcatctct
     1801 gttccatgtt tctaatgatt actcattcat tcgatattac acaaggaccc tttaccaatt
     1861 ccagtttcaa gaagcacttt gtcaagcagc taaacatgaa ggccctctgc acaaatgtga
     1921 catctcaaac tctacagaag ctggacagaa actgttcaat atgctgaggc ttggaaaatc
     1981 agaaccctgg accctagcat tggaaaatgt tgtaggagca aagaacatga atgtaaggcc
     2041 actgctcaac tactttgagc ccttatttac ctggctgaaa gaccagaaca agaattcttt
     2101 tgtgggatgg agtaccgact ggagtccata tgcagacgga tccggtggag gatcttgcca
     2161 tccaagactt agtctccatc gcccagctct tgaagatttg ttgctgggaa gcgaggccaa
     2221 tctcacctgc acactgactg gccttagaga cgcatccggc gtcacgttca catggacgcc
     2281 tagttcagga aaaagcgccg tacaggggcc acctgaaagg gacctgtgtg ggtgctattc
     2341 agttagttct gtcctgcctg gatgcgcgga accgtggaat catgggaaaa cttttacctg
     2401 cacggcggct tatccagaaa gcaaaacccc tctcacggcg acccttagca agagcggaaa
     2461 tacttttcgg ccagaagttc acttgttgcc tccgccgagc gaggaattgg cacttaatga
     2521 gctcgttacc ctcacttgcc ttgcacgagg gttctctcct aaggatgtgc tcgttcgatg
     2581 gttgcagggc tcacaggaac ttcctaggga gaaatacttg acctgggcct ctagacagga
     2641 accgtctcaa ggcaccacaa catttgcggt tacaagtatc ttgcgcgtag cagctgagga
     2701 ctggaagaag ggtgacacct tttcctgtat ggttggacat gaggccctgc cacttgcatt
     2761 tacccagaaa actatagata ggcttgctgg gaagcccacg cacgtcaacg tcagtgtggt
     2821 aatggccgaa gtggatggta catgttactg ataactcgag tctagagggc ccgtttaaac
     2881 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     2941 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     3001 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     3061 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     3121 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     3181 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     3241 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     3301 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     3361 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     3421 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     3481 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     3541 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     3601 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     3661 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     3721 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     3781 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     3841 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     3901 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     3961 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     4021 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     4081 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     4141 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     4201 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     4261 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     4321 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     4381 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     4441 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     4501 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     4561 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     4621 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     4681 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     4741 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     4801 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     4861 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     4921 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     4981 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     5041 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     5101 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     5161 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     5221 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     5281 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     5341 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     5401 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     5461 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     5521 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     5581 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     5641 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     5701 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     5761 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     5821 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     5881 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     5941 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     6001 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     6061 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     6121 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     6181 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     6241 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     6301 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     6361 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     6421 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     6481 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     6541 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     6601 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     6661 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     6721 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     6781 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     6841 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     6901 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     6961 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     7021 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     7081 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     7141 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     7201 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     7261 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     7321 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     7381 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     7441 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     7501 gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga ctagttatta
     7561 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
     7621 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
     7681 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
     7741 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
     7801 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
     7861 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatg
//
