LOCUS       sequence-343861-        5170 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  For making AAV9.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5170)
  AUTHORS   Akerberg BN, Gu F, VanDusen NJ, Zhang X, Dong R, Li K, Zhang B, Zhou
            B, Sethi I, Ma Q, Wasson L, Wen T, Liu J, Dong K, Conlon FL, Zhou J,
            Yuan GC, Zhou P, Pu WT
  TITLE     A reference map of murine cardiac transcription factor chromatin 
            occupancy identifies dynamic and conserved enhancers.
  JOURNAL   Nat Commun. 2019 Oct 28;10(1):4907. doi: 10.1038/s41467-019-12812-3.
  PUBMED    31659164
REFERENCE   2  (bases 1 to 5170)
  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: "Nat 
            Commun."; date: "2019-10-28"; volume: "10(1):4907. doi"; pages: " 
            10.1038/s41467-019-12812-3"
COMMENT     Sequence Label: penaav-tnt-3xha-p2a-gfp
FEATURES             Location/Qualifiers
     source          1..5170
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     intron          593..725
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     708..727
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        770..788
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             843..932
                     /codon_start=1
                     /product="three tandem HA epitope tags"
                     /label=3xHA
                     /translation="YPYDVPDYAGYPYDVPDYAGSYPYDVPDYA"
     CDS             975..1031
                     /codon_start=1
                     /product="2A peptide from porcine teschovirus-1 
                     polyprotein"
                     /label=P2A
                     /note="Eukaryotic ribosomes fail to insert a peptide bond 
                     between the Gly and Pro residues, yielding separate 
                     polypeptides."
                     /translation="ATNFSLLKQAGDVEENPGP"
     CDS             join(1035..1037,1038..1040,1041..1754)
                     /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(1035..1037,1038..1040,1041..1751)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1080..1101)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1341..1360)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1688..1709
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1828..1847)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1893..1948
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2041..2170
                     /label=AAV2 ITR
     primer_bind     complement(2189..2206)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2189..2205)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2198..2220)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2347..2802
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2434..2453)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2644..2665
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2828..2932
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2933..3001,3002..3793)
                     /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(3151..3170)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3964..4552
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4453..4472
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    4840..4861
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4876..4881,4882..4899,4900..4906)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4914..4930
                     /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)."
     primer_bind     4919..4941
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4938..4954
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4938..4954
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   4992..5121
                     /label=AAV2 ITR
ORIGIN
        1 gatcctctag aactatagct agaattcgcc cttacgggcc ccccctcgag gtcgggataa
       61 aagcagtctg ggctttcaca tgacagcatc tggggctgcg gcagagggtc gggtccgaag
      121 cgctgcctta tcagcgtccc cagccctggg aggtgacagc tggctggctt gtgtcagccc
      181 ctcgggcact cacgtatctc cgtccgacgg gtttaaaata gcaaaactct gaggccacac
      241 aatagcttgg gcttatatgg gctcctgtgg gggaaggggg agcacggagg gggccggggc
      301 cgctgctgcc aaaatagcag ctcacaagtg ttgcattcct ctctgggcgc cgggcacatt
      361 cctgctggct ctgcccgccc cggggtgggc gccgggggga ccttaaagcc tctgcccccc
      421 aaggagccct tcccagacag ccgccggcac ccaccgctcc gtgggacgat ccccgaagct
      481 ctagagcttt attgcggtag tttatcacag ttaaattgct aacgcagtca gtgcttctga
      541 cacaacagtc tcgaacttaa gctgcagaag ttggtcgtga ggcactgggc aggtaagtat
      601 caaggttaca agacaggttt aaggagacca atagaaactg ggcttgtcga gacagagaag
      661 actcttgcgt ttctgatagg cacctattgg tcttactgac atccactttg cctttctctc
      721 cacaggtgtc cactcccagt tcaattacag ctcttaaggc tagagtactt aatacgactc
      781 actataggct agcctcgaga agcggccgca ctactccgcg gactactact agtatggccg
      841 tttacccata cgatgttcct gactatgcgg gctatcccta tgacgtcccg gactatgcag
      901 gatcctatcc atatgacgtt ccagattacg ctaccggtgg aaagcttgga caattgggag
      961 agctcggatc cggagccacg aacttctctc tgttaaagca agcaggagac gtggaagaaa
     1021 accccggtcc tgccatggtg agcaagggcg aggagctgtt caccggggtg gtgcccatcc
     1081 tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc gagggcgagg
     1141 gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc aagctgcccg
     1201 tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc agccgctacc
     1261 ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc tacgtccagg
     1321 agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag gtgaagttcg
     1381 agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag gaggacggca
     1441 acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat atcatggccg
     1501 acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc gaggacggca
     1561 gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc cccgtgctgc
     1621 tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc aacgagaagc
     1681 gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc ggcatggacg
     1741 agctgtacaa gtaataagct cgcgtggtac ctctagagtc gacccgggcg gcctcgagga
     1801 cggggtgaac tacgcctgag gatccgatct ttttccctct gccaaaaatt atggggacat
     1861 catgaagccc cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat
     1921 agtgtgttgg aattttttgt gtctctcact cggaagcaat tcgttgatct gaatttcgac
     1981 cacccataat acccattacc ctggtagata agtagcatgg cgggttaatc attaactaca
     2041 aggaacccct agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg
     2101 ccgggcgacc aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc
     2161 gagcgcgcag ccttaattaa cctaattcac tggccgtcgt tttacaacgt cgtgactggg
     2221 aaaaccctgg cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc
     2281 gtaatagcga agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg
     2341 aatgggacgc gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg
     2401 tgaccgctac acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc
     2461 tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc
     2521 gatttagtgc tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta
     2581 gtgggccatc gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta
     2641 atagtggact cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg
     2701 atttataagg gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa
     2761 aatttaacgc gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg
     2821 aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct
     2881 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     2941 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     3001 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     3061 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     3121 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga
     3181 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     3241 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     3301 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     3361 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     3421 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc
     3481 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     3541 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     3601 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     3661 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     3721 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     3781 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     3841 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     3901 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     3961 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     4021 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     4081 cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca
     4141 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     4201 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     4261 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     4321 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     4381 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     4441 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     4501 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     4561 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     4621 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     4681 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc
     4741 aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag
     4801 gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca
     4861 ttaggcaccc caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag
     4921 cggataacaa tttcacacag gaaacagcta tgaccatgat tacgccagat ttaattaagg
     4981 ccttaattag gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
     5041 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
     5101 actccatcac taggggttcc ttgtagttaa tgattaaccc gccatgctac ttatctacca
     5161 gggtaatggg
//
