LOCUS       sequence-292623-        7891 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression plasmid for soluble ACE2-Fc fusion (IgG1) high 
            affinity 4.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7891)
  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 7891)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; doi: 
            "10.1126/science.abc0870"; journalName: "Science"; date: "2020-09-4-
            4"; volume: "369"; issue: "6508"; pages: "1261-1265"
COMMENT     Sequence Label: pcDNA3-sACE2v4-Fc(IgG1)
FEATURES             Location/Qualifiers
     source          1..7891
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        135..514
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        515..718
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     669..689
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     763..782
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        763..781
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3382..3399)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3388..3612
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3658..4086
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3745..3764)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3955..3976
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4095..4115)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4100..4429
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4280..4415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4342..4361
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             4496..5290
                     /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(4550..4569)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5160..5179
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5464..5585
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5501..5520)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5555..5574
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5634..5650)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5634..5650)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5647..5669)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5658..5674
                     /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(join(5682..5688,5689..5706,5707..5712))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5727..5748
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5865..5882)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6036..6624)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6116..6135)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6795..7586,7587..7655))
                     /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     7418..7437
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7656..7760)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7835..7854)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 ggaggtcgct gagtagtgcg cgagcaaaat ttaagctaca acaaggcaag gcttgaccga
       61 caattgcatg aagaatctgc ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc
      121 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc
      181 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc
      241 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt
      301 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca
      361 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg
      421 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca
      481 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa
      541 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa
      601 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc
      661 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct
      721 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag
      781 ggagacccaa gctggctagc gccaccatgt caagctcttc ctggctcctt ctcagccttg
      841 ttgctgtaac tgctgctcag tccaccattg aggaacaggc caagacattt ttggacaagt
      901 ttaacgccga agccgaagac ctgttctatc aaagttcact tgcttcttgg aattataaca
      961 ccaatattac tgaagagaat gtccaaaaca tgaataatgc tggggacaaa tggtctgcct
     1021 ttttaaagga acagtccaca actgcccaaa tgtatccact acaagaaatt cagaatctca
     1081 cagtcaagct tcagctgcag gctcttcagc aaaatgggtc ttcagtgctc tcagaagaca
     1141 agagcaaacg gttgaacaca attctaaata caatgagcac catctacagt actggaaaag
     1201 tttgtaaccc agataatcca caagaatgct tattacttga accaggtttg aatgaaataa
     1261 tggcaaacag tttagactac aatgagaggc tctgggcttg ggaaagctgg agatctgagg
     1321 tcggcaagca gctgaggcca ttatatgaag agtatgtggt cttgaaaaat gagatggcaa
     1381 gagcaaatca ttatgaggac tatggggatt attggagagg agactatgaa gtaaatgggg
     1441 tagatggcta tgactacagc cgcggccagt tgattgaaga tgtggaacat acctttgaag
     1501 agattaaacc attatatgaa catcttcatg cctatgtgag ggcaaagttg atgaatgcct
     1561 atccttccta tatcagtcca attggatgcc tccctgctca tttgcttggt gatatgtggg
     1621 gtagattttg gacaaatctg tactctttga cagttccctt tggacagaaa ccaaacatag
     1681 atgttactga tgcaatggtg gaccaggcct gggatgcaca gagaatattc aaggaggccg
     1741 agaagttctt tgtatctgtt ggtcttccta atatgactca aggattctgg gaatattcca
     1801 tgctaacgga cccaggaaat gttcagaaag cagtctgcca tcccacagct tgggacctgg
     1861 ggaagggcga cttcaggatt cttatgtgca caaaggtgac aatggacgac ttcctgacag
     1921 ctcatcatga gatggggcat atccagtatg atatggcata tcttgcacaa ccttttctgc
     1981 taagaaatgg agctaatgaa ggattccatg aagctgttgg ggaaatcatg tcactttctg
     2041 cagccacacc taagcattta aaatccattg gtcttctgtc acccgatttt caagaagaca
     2101 atgaaacaga aataaacttc ctgctcaaac aagcactcac gattgttggg actctgccat
     2161 ttacttacat gttagagaag tggaggtgga tggtctttaa aggggaaatt cccaaagacc
     2221 agtggatgaa aaagtggtgg gagatgaagc gagagatagt tggggtggtg gaacctgtgc
     2281 cccatgatga aacatactgt gaccccgcat ctctgttcca tgtttctaat gattactcat
     2341 tcattcgata ttacacaagg accctttacc aattccagtt tcaagaagca ctttgtcaag
     2401 cagctaaaca tgaaggccct ctgcacaaat gtgacatctc aaactctaca gaagctggac
     2461 agaaactgtt caatatgctg aggcttggaa aatcagaacc ctggacccta gcattggaaa
     2521 atgttgtagg agcaaagaac atgaatgtaa ggccactgct caactacttt gagcccttat
     2581 ttacctggct gaaagaccag aacaagaatt cttttgtggg atggagtacc gactggagtc
     2641 catatgcaga cggatccgac aaaactcaca catgcccacc gtgcccagca cctgaactcc
     2701 tggggggacc gtcagtcttc ctcttccccc caaaacccaa ggacaccctc atgatctccc
     2761 ggacccctga ggtcacatgc gtggtggtgg acgtgagcca cgaagaccct gaggtcaagt
     2821 tcaactggta cgtggacggc gtggaggtgc ataatgccaa gacaaagccg cgggaggagc
     2881 agtacaacag cacgtaccgt gtggtcagcg tcctcaccgt cctgcaccag gactggctga
     2941 atggcaagga gtacaagtgc aaggtctcca acaaagccct cccagccccc atcgagaaaa
     3001 ccatctccaa agccaaaggg cagccccgag aaccacaggt gtacaccctg cccccatccc
     3061 gggaggagat gaccaagaac caggtcagcc tgacctgcct ggtcaaaggc ttctatccca
     3121 gcgacatcgc cgtggagtgg gagagcaatg ggcagccgga gaacaactac aagaccacgc
     3181 ctcccgtgct ggactccgac ggctccttct tcctctacag caagctcacc gtggacaaga
     3241 gcaggtggca gcaggggaac gtcttctcat gctccgtgat gcacgaggct ctgcacaacc
     3301 actacacgca gaagagcctc tccctgtctc cgggtaaatg ataactcgag tctagagggc
     3361 ccgtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt
     3421 gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat
     3481 aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg
     3541 tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg
     3601 tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg
     3661 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     3721 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     3781 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     3841 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     3901 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     3961 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     4021 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     4081 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     4141 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     4201 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     4261 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     4321 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     4381 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     4441 agcttgtata tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat
     4501 tgaacaagat ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta
     4561 tgactgggca caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca
     4621 ggggcgcccg gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga
     4681 cgaggcagcg cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga
     4741 cgttgtcact gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct
     4801 cctgtcatct caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg
     4861 gctgcatacg cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga
     4921 gcgagcacgt actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca
     4981 tcaggggctc gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga
     5041 ggatctcgtc gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg
     5101 cttttctgga ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc
     5161 gttggctacc cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt
     5221 gctttacggt atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga
     5281 gttcttctga gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca
     5341 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     5401 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     5461 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     5521 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     5581 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     5641 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     5701 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     5761 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     5821 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     5881 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5941 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     6001 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6061 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6121 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6181 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6241 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6301 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6361 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6421 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6481 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6541 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6601 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6661 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6721 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6781 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6841 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6901 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6961 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     7021 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7081 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7141 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7201 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7261 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7321 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7381 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7441 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7501 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7561 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7621 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7681 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7741 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     7801 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     7861 taagccagta tctgctccct gcttgtgtgt t
//
