LOCUS       sequence-310391-        7834 bp    DNA     circular SYN 14-APR-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  JOURNAL   Unpublished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 Apr 14, 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            "Unpublished"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   1..130
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        231..1409
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          462..1400
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1357..1377
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     CDS             3860..3889
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3908..3931
                     /codon_start=1
                     /product="FLAG(R) epitope tag, followed by an enterokinase 
                     cleavage site"
                     /label=FLAG
                     /translation="DYKDDDDK"
     misc_feature    3961..4549
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(4014..4034)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(4432..4443)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    4581..5057
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(4780..4799)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   5097..5237
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   5097..5226
                     /label=AAV2 ITR
     rep_origin      5312..5767
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5399..5418)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5609..5630
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5784..5803)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5903..5925
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5963..5981)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6049..6153
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6154..6222,6223..7014)
                     /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(6372..6391)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7185..7773
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7674..7693
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct gcggccgcac gcgtaagctt tgcaaagatg gataaagttt taaacagaga
      181 ggaatctttg cagctaatgg accttctagg tcttgaaagg agtgggaatt ggctccggtg
      241 cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg ggaggggtcg
      301 gcaattgaac cggtgcctag agaaggtggc gcggggtaaa ctgggaaagt gatgtcgtgt
      361 actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca gtagtcgccg
      421 tgaacgttct ttttcgcaac gggtttgccg ccagaacaca ggtaagtgcc gtgtgtggtt
      481 cccgcgggcc tggcctcttt acgggttatg gcccttgcgt gccttgaatt acttccacct
      541 ggctgcagta cgtgattctt gatcccgagc ttcgggttgg aagtgggtgg gagagttcga
      601 ggccttgcgc ttaaggagcc ccttcgcctc gtgcttgagt tgaggcctgg cctgggcgct
      661 ggggccgccg cgtgcgaatc tggtggcacc ttcgcgcctg tctcgctgct ttcgataagt
      721 ctctagccat ttaaaatttt tgatgacctg ctgcgacgct ttttttctgg caagatagtc
      781 ttgtaaatgc gggccaagat ctgcacactg gtatttcggt ttttggggcc gcgggcggcg
      841 acggggcccg tgcgtcccag cgcacatgtt cggcgaggcg gggcctgcga gcgcggccac
      901 cgagaatcgg acgggggtag tctcaagctg gccggcctgc tctggtgcct ggcctcgcgc
      961 cgccgtgtat cgccccgccc tgggcggcaa ggctggcccg gtcggcacca gttgcgtgag
     1021 cggaaagatg gccgcttccc ggccctgctg cagggagctc aaaatggagg acgcggcgct
     1081 cgggagagcg ggcgggtgag tcacccacac aaaggaaaag ggcctttccg tcctcagccg
     1141 tcgcttcatg tgactccacg gagtaccggg cgccgtccag gcacctcgat tagttctcga
     1201 gcttttggag tacgtcgtct ttaggttggg gggaggggtt ttatgcgatg gagtttcccc
     1261 acactgagtg ggtggagact gaagttaggc cagcttggca cttgatgtaa ttctccttgg
     1321 aatttgccct ttttgagttt ggatcttggt tcattctcaa gcctcagaca gtggttcaaa
     1381 gtttttttct tccatttcag gtgtcgtgag gtaccggatc gccgccatgt caagctcttc
     1441 ctggctcctt ctcagccttg ttgctgtaac tgctgctcag tccaccattg aggaacaggc
     1501 caagacattt ttggacaagt ttaaccacga agccgaagac ctgttctatc aaagttcact
     1561 tgcttcttgg aattataaca ccaatattac tgaagagaat gtccaaaaca tgaataatgc
     1621 tggggacaaa tggtctgcct ttttaaagga acagtccaca cttgcccaaa tgtatccact
     1681 acaagaaatt cagaatctca cagtcaagct tcagctgcag gctcttcagc aaaatgggtc
     1741 ttcagtgctc tcagaagaca agagcaaacg gttgaacaca attctaaata caatgagcac
     1801 catctacagt actggaaaag tttgtaaccc agataatcca caagaatgct tattacttga
     1861 accaggtttg aatgaaataa tggcaaacag tttagactac aatgagaggc tctgggcttg
     1921 ggaaagctgg agatctgagg tcggcaagca gctgaggcca ttatatgaag agtatgtggt
     1981 cttgaaaaat gagatggcaa gagcaaatca ttatgaggac tatggggatt attggagagg
     2041 agactatgaa gtaaatgggg tagatggcta tgactacagc cgcggccagt tgattgaaga
     2101 tgtggaacat acctttgaag agattaaacc attatatgaa catcttcatg cctatgtgag
     2161 ggcaaagttg atgaatgcct atccttccta tatcagtcca attggatgcc tccctgctca
     2221 tttgcttggt gatatgtggg gtagattttg gacaaatctg tactctttga cagttccctt
     2281 tggacagaaa ccaaacatag atgttactga tgcaatggtg gaccaggcct gggatgcaca
     2341 gagaatattc aaggaggccg agaagttctt tgtatctgtt ggtcttccta atatgactca
     2401 aggattctgg gaaaattcca tgctaacgga cccaggaaat gttcagaaag cagtctgcca
     2461 tcccacagct tgggacctgg ggaagggcga cttcaggatc cttatgtgca caaaggtgac
     2521 aatggacgac ttcctgacag ctcatcatga gatggggcat atccagtatg atatggcata
     2581 tgctgcacaa ccttttctgc taagaaatgg agctaatgaa ggattccatg aagctgttgg
     2641 ggaaatcatg tcactttctg cagccacacc taagcattta aaatccattg gtcttctgtc
     2701 acccgatttt caagaagaca atgaaacaga aataaacttc ctgctcaaac aagcactcac
     2761 gattgttggg actctgccat ttacttacat gttagagaag tggaggtgga tggtctttaa
     2821 aggggaaatt cccaaagacc agtggatgaa aaagtggtgg gagatgaagc gagagatagt
     2881 tggggtggtg gaacctgtgc cccatgatga aacatactgt gaccccgcat ctctgttcca
     2941 tgtttctaat gattactcat tcattcgata ttacacaagg accctttacc aattccagtt
     3001 tcaagaagca ctttgtcaag cagctaaaca tgaaggccct ctgcacaaat gtgacatctc
     3061 aaactctaca gaagctggac agaaactgtt caatatgctg aggcttggaa aatcagaacc
     3121 ctggacccta gcattggaaa atgttgtagg agcaaagaac atgaatgtaa ggccactgct
     3181 caactacttt gagcccttat ttacctggct gaaagaccag aacaagaatt cttttgtggg
     3241 atggagtacc gactggagtc catatgcaga ccaaagcatc aaagtgagga taagcctaaa
     3301 atcagctctt ggagataaag catatgaatg gaacgacaat gaaatgtacc tgttccgatc
     3361 atctgttgca tatgctatga ggcagtactt tttaaaagta aaaaatcaga tgattctttt
     3421 tggggaggag gatgtgcgag tggctaattt gaaaccaaga atctccttta atttctttgt
     3481 cactgcacct aaaaatgtgt ctgatatcat tcctagaact gaagttgaaa aggccatcag
     3541 gatgtcccgg agccgtatca atgatgcttt ccgtctgaat gacaacagcc tagagtttct
     3601 ggggatacag ccaacacttg gacctcctaa ccagccccct gtttccatat ggctgattgt
     3661 ttttggagtt gtgatgggag tgatagtggt tggcattgtc atcctgatct tcactgggat
     3721 cagagatcgg aagaagaaaa ataaagcaag aagtggagaa aatccttatg cctccatcga
     3781 tattagcaaa ggagaaaata atccaggatt ccaaaacact gatgatgttc agacctcctt
     3841 tacgcgtacg cggccgctcg agcagaaact catctcagaa gaggatctgg cagcaaatga
     3901 tatcctggat tacaaggatg acgacgataa ggtttaaaat tcgatatcaa gcttatcgat
     3961 aatcaacctc tggattacaa aatttgtgaa agattgactg gtattcttaa ctatgttgct
     4021 ccttttacgc tatgtggata cgctgcttta atgcctttgt atcatgctat tgcttcccgt
     4081 atggctttca ttttctcctc cttgtataaa tcctggttgc tgtctcttta tgaggagttg
     4141 tggcccgttg tcaggcaacg tggcgtggtg tgcactgtgt ttgctgacgc aacccccact
     4201 ggttggggca ttgccaccac ctgtcagctc ctttccggga ctttcgcttt ccccctccct
     4261 attgccacgg cggaactcat cgccgcctgc cttgcccgct gctggacagg ggctcggctg
     4321 ttgggcactg acaattccgt ggtgttgtcg gggaaatcat cgtcctttcc ttggctgctc
     4381 gcctatgttg ccacctggat tctgcgcggg acgtccttct gctacgtccc ttcggccctc
     4441 aatccagcgg accttccttc ccgcggcctg ctgccggctc tgcggcctct tccgcgtctt
     4501 cgccttcgcc ctcagacgag tcggatctcc ctttgggccg cctccccgca tcgataccga
     4561 gcgctgctcg agagatctac gggtggcatc cctgtgaccc ctccccagtg cctctcctgg
     4621 ccctggaagt tgccactcca gtgcccacca gccttgtcct aataaaatta agttgcatca
     4681 ttttgtctga ctaggtgtcc ttctataata ttatggggtg gaggggggtg gtatggagca
     4741 aggggcaagt tgggaagaca acctgtaggg cctgcggggt ctattgggaa ccaagctgga
     4801 gtgcagtggc acaatcttgg ctcactgcaa tctccgcctc ctgggttcaa gcgattctcc
     4861 tgcctcagcc tcccgagttg ttgggattcc aggcatgcat gaccaggctc agctaatttt
     4921 tgtttttttg gtagagacgg ggtttcacca tattggccag gctggtctcc aactcctaat
     4981 ctcaggtgat ctacccacct tggcctccca aattgctggg attacaggcg tgaaccactg
     5041 ctcccttccc tgtccttctg attttgtagg taaccacgtg cggaccgagc ggccgcagga
     5101 acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca ctgaggccgg
     5161 gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga gcgagcgagc
     5221 gcgcagctgc ctgcaggggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     5281 ttcacaccgc atacgtcaaa gcaaccatag tacgcgccct gtagcggcgc attaagcgcg
     5341 gcgggtgtgg tggttacgcg cagcgtgacc gctacacttg ccagcgcctt agcgcccgct
     5401 cctttcgctt tcttcccttc ctttctcgcc acgttcgccg gctttccccg tcaagctcta
     5461 aatcgggggc tccctttagg gttccgattt agtgctttac ggcacctcga ccccaaaaaa
     5521 cttgatttgg gtgatggttc acgtagtggg ccatcgccct gatagacggt ttttcgccct
     5581 ttgacgttgg agtccacgtt ctttaatagt ggactcttgt tccaaactgg aacaacactc
     5641 aactctatct cgggctattc ttttgattta taagggattt tgccgatttc ggtctattgg
     5701 ttaaaaaatg agctgattta acaaaaattt aacgcgaatt ttaacaaaat attaacgttt
     5761 acaattttat ggtgcactct cagtacaatc tgctctgatg ccgcatagtt aagccagccc
     5821 cgacacccgc caacacccgc tgacgcgccc tgacgggctt gtctgctccc ggcatccgct
     5881 tacagacaag ctgtgaccgt ctccgggagc tgcatgtgtc agaggttttc accgtcatca
     5941 ccgaaacgcg cgagacgaaa gggcctcgtg atacgcctat ttttataggt taatgtcatg
     6001 ataataatgg tttcttagac gtcaggtggc acttttcggg gaaatgtgcg cggaacccct
     6061 atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga
     6121 taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc
     6181 cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg
     6241 aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc
     6301 aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact
     6361 tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc
     6421 ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag
     6481 catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat
     6541 aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt
     6601 ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa
     6661 gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc
     6721 aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg
     6781 gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt
     6841 gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca
     6901 gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat
     6961 gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca
     7021 gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg
     7081 atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg
     7141 ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt
     7201 ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg
     7261 ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata
     7321 ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca
     7381 ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag
     7441 tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc
     7501 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
     7561 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
     7621 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
     7681 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
     7741 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
     7801 ttcctggcct tttgctggcc ttttgctcac atgt
//
