LOCUS       sequence-174972-        6972 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Recombinant single-stranded AAV transfer vector expressing Firefly 
            Luciferase under the EF1a promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6972)
  AUTHORS   Paulk NK, Pekrun K, Zhu E, Nygaard S, Li B, Xu J, Chu K, Leborgne C,
            Dane AP, Haft A, Zhang Y, Zhang F, Morton C, Valentine MB, Davidoff 
            AM, Nathwani AC, Mingozzi F, Grompe M, Alexander IE, Lisowski L, Kay
            MA
  TITLE     Bioengineered AAV Capsids with Combined High Human Liver 
            Transduction In Vivo and Unique Humoral Seroreactivity.
  JOURNAL   Mol Ther. 2017 Sep 25. pii: S1525-0016(17)30437-9. doi: 
            10.1016/j.ymthe.2017.09.021.
  PUBMED    29055620
REFERENCE   2  (bases 1 to 6972)
  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: "Mol Ther. 
            2017 Sep 25. pii: S1525-0016(17)30437-9. doi: 
            10.1016/j.ymthe.2017.09.021."
COMMENT     Sequence Label: pAAV-EF1a-FLuc-WPRE-HGHpA
FEATURES             Location/Qualifiers
     source          1..6972
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   26..155
                     /label=AAV2 ITR (alternate)
                     /note="Functional equivalent of wild-type AAV2 ITR"
     promoter        256..1434
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          487..1425
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     1382..1402
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     regulatory      1441..1450
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             1447..3099
                     /codon_start=1
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="synthetic luc2 version of the luciferase gene"
                     /translation="MEDAKNIRKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGETLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     misc_feature    3124..3712
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3177..3197)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(3595..3606)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     polyA_signal    3744..4220
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     primer_bind     complement(3943..3962)
                     /label=hGH-PA-R
                     /note="Human growth hormone terminator, reverse primer"
     repeat_region   4260..4400
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4260..4389
                     /label=AAV2 ITR
     rep_origin      4475..4930
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4562..4581)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4772..4793
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4947..4966)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5066..5088
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5126..5144)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5212..5316
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5317..5385,5386..6177)
                     /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(5535..5554)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6348..6936
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6837..6856
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 ttttgctggc cttttgctca catgtcctgc aggcagctgc gcgctcgctc gctcactgag
       61 gccgcccggg cgtcgggcga cctttggtcg cccggcctca gtgagcgagc gagcgcgcag
      121 agagggagtg gccaactcca tcactagggg ttcctgcggc cgcacgcgta agctttgcaa
      181 agatggataa agttttaaac agagaggaat ctttgcagct aatggacctt ctaggtcttg
      241 aaaggagtgg gaattggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
      301 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      361 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      421 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      481 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
      541 tgcgtgcctt gaattacttc cacctggctg cagtacgtga ttcttgatcc cgagcttcgg
      601 gttggaagtg ggtgggagag ttcgaggcct tgcgcttaag gagccccttc gcctcgtgct
      661 tgagttgagg cctggcctgg gcgctggggc cgccgcgtgc gaatctggtg gcaccttcgc
      721 gcctgtctcg ctgctttcga taagtctcta gccatttaaa atttttgatg acctgctgcg
      781 acgctttttt tctggcaaga tagtcttgta aatgcgggcc aagatctgca cactggtatt
      841 tcggtttttg gggccgcggg cggcgacggg gcccgtgcgt cccagcgcac atgttcggcg
      901 aggcggggcc tgcgagcgcg gccaccgaga atcggacggg ggtagtctca agctggccgg
      961 cctgctctgg tgcctggcct cgcgccgccg tgtatcgccc cgccctgggc ggcaaggctg
     1021 gcccggtcgg caccagttgc gtgagcggaa agatggccgc ttcccggccc tgctgcaggg
     1081 agctcaaaat ggaggacgcg gcgctcggga gagcgggcgg gtgagtcacc cacacaaagg
     1141 aaaagggcct ttccgtcctc agccgtcgct tcatgtgact ccacggagta ccgggcgccg
     1201 tccaggcacc tcgattagtt ctcgagcttt tggagtacgt cgtctttagg ttggggggag
     1261 gggttttatg cgatggagtt tccccacact gagtgggtgg agactgaagt taggccagct
     1321 tggcacttga tgtaattctc cttggaattt gccctttttg agtttggatc ttggttcatt
     1381 ctcaagcctc agacagtggt tcaaagtttt tttcttccat ttcaggtgtc gtgaggtacc
     1441 gccaccatgg aagatgccaa aaacattagg aagggcccag cgccattcta cccactcgaa
     1501 gacgggaccg ccggcgagca gctgcacaaa gccatgaagc gctacgccct ggtgcccggc
     1561 accatcgcct ttaccgacgc acatatcgag gtggacatta cctacgccga gtacttcgag
     1621 atgagcgttc ggctggcaga agctatgaag cgctatgggc tgaatacaaa ccatcggatc
     1681 gtggtgtgca gcgagaatag cttgcagttc ttcatgcccg tgttgggtgc cctgttcatc
     1741 ggtgtggctg tggccccagc taacgacatc tacaacgagc gcgagctgct gaacagcatg
     1801 ggcatcagcc agcccaccgt cgtattcgtg agcaagaaag ggctgcaaaa gatcctcaac
     1861 gtgcaaaaga agctaccgat catacaaaag atcatcatca tggatagcaa gaccgactac
     1921 cagggcttcc aaagcatgta caccttcgtg acttcccatt tgccacccgg cttcaacgag
     1981 tacgacttcg tgcccgagag cttcgaccgg gacaaaacca tcgccctgat catgaacagt
     2041 agtggcagta ccggattgcc caagggcgta gccctaccgc accgcaccgc ttgtgtccga
     2101 ttcagtcatg cccgcgaccc catcttcggc aaccagatca tccccgacac cgctatcctc
     2161 agcgtggtgc catttcacca cggcttcggc atgttcacca cgctgggcta cttgatctgc
     2221 ggctttcggg tcgtgctcat gtaccgcttc gaggaggagc tattcttgcg cagcttgcaa
     2281 gactataaga ttcaatctgc cctgctggtg cccacactat ttagcttctt cgctaagagc
     2341 actctcatcg acaagtacga cctaagcaac ttgcacgaga tcgccagcgg cggggcgccg
     2401 ctcagcaagg aggtaggtga ggccgtggcc aaacgcttcc acctaccagg catccgccag
     2461 ggctacggcc tgacagaaac aaccagcgcc attctgatca cccccgaagg ggacgacaag
     2521 cctggcgcag taggcaaggt ggtgcccttc ttcgaggcta aggtggtgga cttggacacc
     2581 ggtgagacac tgggtgtgaa ccagcgcggc gagctgtgcg tccgtggccc catgatcatg
     2641 agcggctacg ttaacaaccc cgaggctaca aacgctctca tcgacaagga cggctggctg
     2701 cacagcggcg acatcgccta ctgggacgag gacgagcact tcttcatcgt ggaccggctg
     2761 aagagcctga tcaaatacaa gggctaccag gtagccccag ccgaactgga gagcatcctg
     2821 ctgcaacacc ccaacatctt cgacgccggg gtcgccggcc tgcccgacga cgatgccggc
     2881 gagctgcccg ccgcagtcgt cgtgctggaa cacggtaaaa ccatgaccga gaaggagatc
     2941 gtggactatg tggccagcca ggttacaacc gccaagaagc tgcgcggtgg tgttgtgttc
     3001 gtggacgagg tgcctaaagg actgaccggc aagttggacg cccgcaagat ccgcgagatt
     3061 ctcattaagg ccaagaaggg cggcaagatc gccgtgtaag aattcgatat caagcttatc
     3121 gataatcaac ctctggatta caaaatttgt gaaagattga ctggtattct taactatgtt
     3181 gctcctttta cgctatgtgg atacgctgct ttaatgcctt tgtatcatgc tattgcttcc
     3241 cgtatggctt tcattttctc ctccttgtat aaatcctggt tgctgtctct ttatgaggag
     3301 ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc
     3361 actggttggg gcattgccac cacctgtcag ctcctttccg ggactttcgc tttccccctc
     3421 cctattgcca cggcggaact catcgccgcc tgccttgccc gctgctggac aggggctcgg
     3481 ctgttgggca ctgacaattc cgtggtgttg tcggggaaat catcgtcctt tccttggctg
     3541 ctcgcctatg ttgccacctg gattctgcgc gggacgtcct tctgctacgt cccttcggcc
     3601 ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt
     3661 cttcgccttc gccctcagac gagtcggatc tccctttggg ccgcctcccc gcatcgatac
     3721 cgagcgctgc tcgagagatc tacgggtggc atccctgtga cccctcccca gtgcctctcc
     3781 tggccctgga agttgccact ccagtgccca ccagccttgt cctaataaaa ttaagttgca
     3841 tcattttgtc tgactaggtg tccttctata atattatggg gtggaggggg gtggtatgga
     3901 gcaaggggca agttgggaag acaacctgta gggcctgcgg ggtctattgg gaaccaagct
     3961 ggagtgcagt ggcacaatct tggctcactg caatctccgc ctcctgggtt caagcgattc
     4021 tcctgcctca gcctcccgag ttgttgggat tccaggcatg catgaccagg ctcagctaat
     4081 ttttgttttt ttggtagaga cggggtttca ccatattggc caggctggtc tccaactcct
     4141 aatctcaggt gatctaccca ccttggcctc ccaaattgct gggattacag gcgtgaacca
     4201 ctgctccctt ccctgtcctt ctgattttgt aggtaaccac gtgcggaccg agcggccgca
     4261 ggaaccccta gtgatggagt tggccactcc ctctctgcgc gctcgctcgc tcactgaggc
     4321 cgggcgacca aaggtcgccc gacgcccggg ctttgcccgg gcggcctcag tgagcgagcg
     4381 agcgcgcagc tgcctgcagg ggcgcctgat gcggtatttt ctccttacgc atctgtgcgg
     4441 tatttcacac cgcatacgtc aaagcaacca tagtacgcgc cctgtagcgg cgcattaagc
     4501 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cttagcgccc
     4561 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     4621 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     4681 aaacttgatt tgggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     4741 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     4801 ctcaactcta tctcgggcta ttcttttgat ttataaggga ttttgccgat ttcggtctat
     4861 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     4921 tttacaattt tatggtgcac tctcagtaca atctgctctg atgccgcata gttaagccag
     4981 ccccgacacc cgccaacacc cgctgacgcg ccctgacggg cttgtctgct cccggcatcc
     5041 gcttacagac aagctgtgac cgtctccggg agctgcatgt gtcagaggtt ttcaccgtca
     5101 tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc tatttttata ggttaatgtc
     5161 atgataataa tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc
     5221 cctatttgtt tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc
     5281 tgataaatgc ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc
     5341 gcccttattc ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg
     5401 gtgaaagtaa aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat
     5461 ctcaacagcg gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc
     5521 acttttaaag ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa
     5581 ctcggtcgcc gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa
     5641 aagcatctta cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt
     5701 gataacactg cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct
     5761 tttttgcaca acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat
     5821 gaagccatac caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg
     5881 cgcaaactat taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg
     5941 atggaggcgg ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt
     6001 attgctgata aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg
     6061 ccagatggta agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg
     6121 gatgaacgaa atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg
     6181 tcagaccaag tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa
     6241 aggatctagg tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt
     6301 tcgttccact gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt
     6361 tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt
     6421 ttgccggatc aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag
     6481 ataccaaata ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta
     6541 gcaccgccta catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat
     6601 aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg
     6661 ggctgaacgg ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg
     6721 agatacctac agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac
     6781 aggtatccgg taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga
     6841 aacgcctggt atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt
     6901 ttgtgatgct cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta
     6961 cggttcctgg cc
//
