LOCUS       sequence-306701-        7834 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Used for expression of human ACE2 with AAV packaging.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7834)
  TITLE     Mouse model of SARS-CoV-2
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 7834)
  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: 
            "Unpublished"
COMMENT     Sequence Label: AAV-hACE2-cMYC-Flag
FEATURES             Location/Qualifiers
     source          1..7834
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   13..142
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        243..1421
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          474..1412
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1369..1389
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3872..3901
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3920..3943
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3973..4561
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4026..4046)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4444..4455)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4593..5069
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4792..4811)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5109..5249
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5109..5238
                     /label=AAV2 ITR
     rep_origin      5324..5779
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5411..5430)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5621..5642
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5796..5815)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5915..5937
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5975..5993)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6061..6165
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6166..6234,6235..7026)
                     /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     complement(6384..6403)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7197..7785
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7686..7705
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttgctcacat gtcctgcagg cagctgcgcg ctcgctcgct cactgaggcc gcccgggcgt
       61 cgggcgacct ttggtcgccc ggcctcagtg agcgagcgag cgcgcagaga gggagtggcc
      121 aactccatca ctaggggttc ctgcggccgc acgcgtaagc tttgcaaaga tggataaagt
      181 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      241 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      301 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      361 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      421 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      481 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      541 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      601 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      661 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      721 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      781 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      841 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      901 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      961 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
     1021 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
     1081 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1141 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1201 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1261 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1321 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1381 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggtaccgga tcgccgccat
     1441 gtcaagctct tcctggctcc ttctcagcct tgttgctgta actgctgctc agtccaccat
     1501 tgaggaacag gccaagacat ttttggacaa gtttaaccac gaagccgaag acctgttcta
     1561 tcaaagttca cttgcttctt ggaattataa caccaatatt actgaagaga atgtccaaaa
     1621 catgaataat gctggggaca aatggtctgc ctttttaaag gaacagtcca cacttgccca
     1681 aatgtatcca ctacaagaaa ttcagaatct cacagtcaag cttcagctgc aggctcttca
     1741 gcaaaatggg tcttcagtgc tctcagaaga caagagcaaa cggttgaaca caattctaaa
     1801 tacaatgagc accatctaca gtactggaaa agtttgtaac ccagataatc cacaagaatg
     1861 cttattactt gaaccaggtt tgaatgaaat aatggcaaac agtttagact acaatgagag
     1921 gctctgggct tgggaaagct ggagatctga ggtcggcaag cagctgaggc cattatatga
     1981 agagtatgtg gtcttgaaaa atgagatggc aagagcaaat cattatgagg actatgggga
     2041 ttattggaga ggagactatg aagtaaatgg ggtagatggc tatgactaca gccgcggcca
     2101 gttgattgaa gatgtggaac atacctttga agagattaaa ccattatatg aacatcttca
     2161 tgcctatgtg agggcaaagt tgatgaatgc ctatccttcc tatatcagtc caattggatg
     2221 cctccctgct catttgcttg gtgatatgtg gggtagattt tggacaaatc tgtactcttt
     2281 gacagttccc tttggacaga aaccaaacat agatgttact gatgcaatgg tggaccaggc
     2341 ctgggatgca cagagaatat tcaaggaggc cgagaagttc tttgtatctg ttggtcttcc
     2401 taatatgact caaggattct gggaaaattc catgctaacg gacccaggaa atgttcagaa
     2461 agcagtctgc catcccacag cttgggacct ggggaagggc gacttcagga tccttatgtg
     2521 cacaaaggtg acaatggacg acttcctgac agctcatcat gagatggggc atatccagta
     2581 tgatatggca tatgctgcac aaccttttct gctaagaaat ggagctaatg aaggattcca
     2641 tgaagctgtt ggggaaatca tgtcactttc tgcagccaca cctaagcatt taaaatccat
     2701 tggtcttctg tcacccgatt ttcaagaaga caatgaaaca gaaataaact tcctgctcaa
     2761 acaagcactc acgattgttg ggactctgcc atttacttac atgttagaga agtggaggtg
     2821 gatggtcttt aaaggggaaa ttcccaaaga ccagtggatg aaaaagtggt gggagatgaa
     2881 gcgagagata gttggggtgg tggaacctgt gccccatgat gaaacatact gtgaccccgc
     2941 atctctgttc catgtttcta atgattactc attcattcga tattacacaa ggacccttta
     3001 ccaattccag tttcaagaag cactttgtca agcagctaaa catgaaggcc ctctgcacaa
     3061 atgtgacatc tcaaactcta cagaagctgg acagaaactg ttcaatatgc tgaggcttgg
     3121 aaaatcagaa ccctggaccc tagcattgga aaatgttgta ggagcaaaga acatgaatgt
     3181 aaggccactg ctcaactact ttgagccctt atttacctgg ctgaaagacc agaacaagaa
     3241 ttcttttgtg ggatggagta ccgactggag tccatatgca gaccaaagca tcaaagtgag
     3301 gataagccta aaatcagctc ttggagataa agcatatgaa tggaacgaca atgaaatgta
     3361 cctgttccga tcatctgttg catatgctat gaggcagtac tttttaaaag taaaaaatca
     3421 gatgattctt tttggggagg aggatgtgcg agtggctaat ttgaaaccaa gaatctcctt
     3481 taatttcttt gtcactgcac ctaaaaatgt gtctgatatc attcctagaa ctgaagttga
     3541 aaaggccatc aggatgtccc ggagccgtat caatgatgct ttccgtctga atgacaacag
     3601 cctagagttt ctggggatac agccaacact tggacctcct aaccagcccc ctgtttccat
     3661 atggctgatt gtttttggag ttgtgatggg agtgatagtg gttggcattg tcatcctgat
     3721 cttcactggg atcagagatc ggaagaagaa aaataaagca agaagtggag aaaatcctta
     3781 tgcctccatc gatattagca aaggagaaaa taatccagga ttccaaaaca ctgatgatgt
     3841 tcagacctcc tttacgcgta cgcggccgct cgagcagaaa ctcatctcag aagaggatct
     3901 ggcagcaaat gatatcctgg attacaagga tgacgacgat aaggtttaaa attcgatatc
     3961 aagcttatcg ataatcaacc tctggattac aaaatttgtg aaagattgac tggtattctt
     4021 aactatgttg ctccttttac gctatgtgga tacgctgctt taatgccttt gtatcatgct
     4081 attgcttccc gtatggcttt cattttctcc tccttgtata aatcctggtt gctgtctctt
     4141 tatgaggagt tgtggcccgt tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac
     4201 gcaaccccca ctggttgggg cattgccacc acctgtcagc tcctttccgg gactttcgct
     4261 ttccccctcc ctattgccac ggcggaactc atcgccgcct gccttgcccg ctgctggaca
     4321 ggggctcggc tgttgggcac tgacaattcc gtggtgttgt cggggaaatc atcgtccttt
     4381 ccttggctgc tcgcctatgt tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc
     4441 ccttcggccc tcaatccagc ggaccttcct tcccgcggcc tgctgccggc tctgcggcct
     4501 cttccgcgtc ttcgccttcg ccctcagacg agtcggatct ccctttgggc cgcctccccg
     4561 catcgatacc gagcgctgct cgagagatct acgggtggca tccctgtgac ccctccccag
     4621 tgcctctcct ggccctggaa gttgccactc cagtgcccac cagccttgtc ctaataaaat
     4681 taagttgcat cattttgtct gactaggtgt ccttctataa tattatgggg tggagggggg
     4741 tggtatggag caaggggcaa gttgggaaga caacctgtag ggcctgcggg gtctattggg
     4801 aaccaagctg gagtgcagtg gcacaatctt ggctcactgc aatctccgcc tcctgggttc
     4861 aagcgattct cctgcctcag cctcccgagt tgttgggatt ccaggcatgc atgaccaggc
     4921 tcagctaatt tttgtttttt tggtagagac ggggtttcac catattggcc aggctggtct
     4981 ccaactccta atctcaggtg atctacccac cttggcctcc caaattgctg ggattacagg
     5041 cgtgaaccac tgctcccttc cctgtccttc tgattttgta ggtaaccacg tgcggaccga
     5101 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     5161 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     5221 gagcgagcga gcgcgcagct gcctgcaggg gcgcctgatg cggtattttc tccttacgca
     5281 tctgtgcggt atttcacacc gcatacgtca aagcaaccat agtacgcgcc ctgtagcggc
     5341 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5401 ttagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     5461 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     5521 gaccccaaaa aacttgattt gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     5581 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     5641 ggaacaacac tcaactctat ctcgggctat tcttttgatt tataagggat tttgccgatt
     5701 tcggtctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     5761 atattaacgt ttacaatttt atggtgcact ctcagtacaa tctgctctga tgccgcatag
     5821 ttaagccagc cccgacaccc gccaacaccc gctgacgcgc cctgacgggc ttgtctgctc
     5881 ccggcatccg cttacagaca agctgtgacc gtctccggga gctgcatgtg tcagaggttt
     5941 tcaccgtcat caccgaaacg cgcgagacga aagggcctcg tgatacgcct atttttatag
     6001 gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg
     6061 cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga
     6121 caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat
     6181 ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca
     6241 gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc
     6301 gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca
     6361 atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg
     6421 caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca
     6481 gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata
     6541 accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag
     6601 ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg
     6661 gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca
     6721 acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta
     6781 atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct
     6841 ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca
     6901 gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag
     6961 gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat
     7021 tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt
     7081 taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa
     7141 cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     7201 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     7261 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     7321 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca ccacttcaag
     7381 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     7441 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     7501 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     7561 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     7621 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     7681 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     7741 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     7801 gcctttttac ggttcctggc cttttgctgg cctt
//
