LOCUS       sequence-305331-        8511 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  pcDNA3.1-ACE2-GFP.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8511)
  AUTHORS   Bartolome A, Liang J, Wang P, Ho DD, Pajvani UB
  TITLE     Angiotensin converting enzyme 2 is a novel target of the 
            gamma-secretase complex.
  JOURNAL   Sci Rep. 2021 May 7;11(1):9803. doi: 10.1038/s41598-021-89379-x.
  PUBMED    33963249
REFERENCE   2  (bases 1 to 8511)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Sci Rep."; 
            date: "2021-05-7"; volume: "11(1):9803. doi"; pages: " 
            10.1038/s41598-021-89379-x"
COMMENT     Sequence Label: pcDNA3.1-ACE2-GFP
FEATURES             Location/Qualifiers
     source          1..8511
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             join(3110..3112,3113..3115,3116..3829)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(3110..3112,3113..3115,3116..3826)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3155..3176)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3416..3435)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3763..3784
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(3870..3887)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3876..4100
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4146..4574
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4233..4252)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4443..4464
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4583..4603)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4588..4917
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4768..4903
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4830..4849
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     complement(5038..5057)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     5646..5665
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    5950..6071
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5987..6006)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6041..6060
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6120..6136)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6120..6136)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6133..6155)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6144..6160
                     /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(6168..6174,6175..6192,6193..6198))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6213..6234
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6351..6368)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6522..7110)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6602..6621)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7281..8072,8073..8141))
                     /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     7904..7923
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(8142..8246)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(8321..8340)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctg
      661 gctagcgttt aaacgggccc tctagaccat gtcaagctct tcctggctcc ttctcagcct
      721 tgttgctgta actgctgctc agtccaccat tgaggaacag gccaagacat ttttggacaa
      781 gtttaaccac gaagccgaag acctgttcta tcaaagttca cttgcttctt ggaattataa
      841 caccaatatt actgaagaga atgtccaaaa catgaataat gctggggaca aatggtctgc
      901 ctttttaaag gaacagtcca cacttgccca aatgtatcca ctacaagaaa ttcagaatct
      961 cacagtcaag cttcagctgc aggctcttca gcaaaatggg tcttcagtgc tctcagaaga
     1021 caagagcaaa cggttgaaca caattctaaa tacaatgagc accatctaca gtactggaaa
     1081 agtttgtaac ccagataatc cacaagaatg cttattactt gaaccaggtt tgaatgaaat
     1141 aatggcaaac agtttagact acaatgagag gctctgggct tgggaaagct ggagatctga
     1201 ggtcggcaag cagctgaggc cattatatga agagtatgtg gtcttgaaaa atgagatggc
     1261 aagagcaaat cattatgagg actatgggga ttattggaga ggagactatg aagtaaatgg
     1321 ggtagatggc tatgactaca gccgcggcca gttgattgaa gatgtggaac atacctttga
     1381 agagattaaa ccattatatg aacatcttca tgcctatgtg agggcaaagt tgatgaatgc
     1441 ctatccttcc tatatcagtc caattggatg cctccctgct catttgcttg gtgatatgtg
     1501 gggtagattt tggacaaatc tgtactcttt gacagttccc tttggacaga aaccaaacat
     1561 agatgttact gatgcaatgg tggaccaggc ctgggatgca cagagaatat tcaaggaggc
     1621 cgagaagttc tttgtatctg ttggtcttcc taatatgact caaggattct gggaaaattc
     1681 catgctaacg gacccaggaa atgttcagaa agcagtctgc catcccacag cttgggacct
     1741 ggggaagggc gacttcagga tccttatgtg cacaaaggtg acaatggacg acttcctgac
     1801 agctcatcat gagatggggc atatccagta tgatatggca tatgctgcac aaccttttct
     1861 gctaagaaat ggagctaatg aaggattcca tgaagctgtt ggggaaatca tgtcactttc
     1921 tgcagccaca cctaagcatt taaaatccat tggtcttctg tcacccgatt ttcaagaaga
     1981 caatgaaaca gaaataaact tcctgctcaa acaagcactc acgattgttg ggactctgcc
     2041 atttacttac atgttagaga agtggaggtg gatggtcttt aaaggggaaa ttcccaaaga
     2101 ccagtggatg aaaaagtggt gggagatgaa gcgagagata gttggggtgg tggaacctgt
     2161 gccccatgat gaaacatact gtgaccccgc atctctgttc catgtttcta atgattactc
     2221 attcattcga tattacacaa ggacccttta ccaattccag tttcaagaag cactttgtca
     2281 agcagctaaa catgaaggcc ctctgcacaa atgtgacatc tcaaactcta cagaagctgg
     2341 acagaaactg ttcaatatgc tgaggcttgg aaaatcagaa ccctggaccc tagcattgga
     2401 aaatgttgta ggagcaaaga acatgaatgt aaggccactg ctcaactact ttgagccctt
     2461 atttacctgg ctgaaagacc agaacaagaa ttcttttgtg ggatggagta ccgactggag
     2521 tccatatgca gaccaaagca tcaaagtgag gataagccta aaatcagctc ttggagataa
     2581 agcatatgaa tggaacgaca atgaaatgta cctgttccga tcatctgttg catatgctat
     2641 gaggcagtac tttttaaaag taaaaaatca gatgattctt tttggggagg aggatgtgcg
     2701 agtggctaat ttgaaaccaa gaatctcctt taatttcttt gtcactgcac ctaaaaatgt
     2761 gtctgatatc attcctagaa ctgaagttga aaaggccatc aggatgtccc ggagccgtat
     2821 caatgatgct ttccgtctga atgacaacag cctagagttt ctggggatac agccaacact
     2881 tggacctcct aaccagcccc ctgtttccat atggctgatt gtttttggag ttgtgatggg
     2941 agtgatagtg gttggcattg tcatcctgat cttcactggg atcagagatc ggaagaagaa
     3001 aaataaagca agaagtggag aaaatcctta tgcctccatc gatattagca aaggagaaaa
     3061 taatccagga ttccaaaaca ctgatgatgt tcagacctcc tttggtacca tggtgagcaa
     3121 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     3181 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3241 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3301 cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3361 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3421 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3481 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3541 caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg gcatcaaggt
     3601 gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3661 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3721 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3781 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaac ttaagcttgg
     3841 atccgcgtta agtttaaacc gctgatcagc ctcgactgtg ccttctagtt gccagccatc
     3901 tgttgtttgc ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct
     3961 ttcctaataa aatgaggaaa ttgcatcgca ttgtctgagt aggtgtcatt ctattctggg
     4021 gggtggggtg gggcaggaca gcaaggggga ggattgggaa gacaatagca ggcatgctgg
     4081 ggatgcggtg ggctctatgg cttctgaggc ggaaagaacc agctggggct ctagggggta
     4141 tccccacgcg ccctgtagcg gcgcattaag cgcggcgggt gtggtggtta cgcgcagcgt
     4201 gaccgctaca cttgccagcg ccctagcgcc cgctcctttc gctttcttcc cttcctttct
     4261 cgccacgttc gccggctttc cccgtcaagc tctaaatcgg gggctccctt tagggttccg
     4321 atttagtgct ttacggcacc tcgaccccaa aaaacttgat tagggtgatg gttcacgtag
     4381 tgggccatcg ccctgataga cggtttttcg ccctttgacg ttggagtcca cgttctttaa
     4441 tagtggactc ttgttccaaa ctggaacaac actcaaccct atctcggtct attcttttga
     4501 tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga tttaacaaaa
     4561 atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa gtccccaggc
     4621 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga
     4681 aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca
     4741 accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat
     4801 tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc
     4861 tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag
     4921 ctcccgggag cttgtatatc cattttcgga tctgatcaag agacaggatg aggatcgttt
     4981 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     5041 ttcggctatg actgggcaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc
     5101 agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact
     5161 gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt
     5221 gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca
     5281 ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat
     5341 gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg
     5401 catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga
     5461 agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga
     5521 cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa
     5581 tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga
     5641 catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt
     5701 cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct
     5761 tgacgagttc ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac
     5821 ctgccatcac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     5881 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     5941 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     6001 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     6061 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     6121 tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc
     6181 ggaagcataa agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg
     6241 ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc
     6301 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     6361 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     6421 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     6481 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     6541 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     6601 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     6661 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     6721 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     6781 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     6841 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6901 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6961 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     7021 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     7081 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     7141 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     7201 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     7261 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     7321 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     7381 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     7441 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     7501 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     7561 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     7621 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     7681 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     7741 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     7801 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     7861 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7921 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7981 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     8041 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     8101 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     8161 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     8221 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac
     8281 ggatcgggag atctcccgat cccctatggt gcactctcag tacaatctgc tctgatgccg
     8341 catagttaag ccagtatctg ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga
     8401 gcaaaattta agctacaaca aggcaaggct tgaccgacaa ttgcatgaag aatctgctta
     8461 gggttaggcg ttttgcgctg cttcgcgatg tacgggccag atatacgcgt t
//
