LOCUS       sequence-454211-        5732 bp    DNA     circular SYN 15-APR-2025
DEFINITION  expressing HA tag and the coding sequence of luciferase by the 
            constitutive CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5732)
  TITLE     Myocardial infarction creates a critical time window for AAV gene 
            therapy
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 5732)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAAV-CMV -HA-Luciferase
FEATURES             Location/Qualifiers
     source          1..5732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     repeat_region   1..130
                     /label=AAV2 ITR
     enhancer        217..596
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        597..800
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     751..771
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             831..857
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             858..2507
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     primer_bind     complement(921..939)
                     /label=LucNrev
                     /note="Firefly luciferase, reverse primer"
     primer_bind     complement(2569..2588)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2634..2689
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2782..2911
                     /label=AAV2 ITR
     primer_bind     complement(2930..2947)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2930..2946)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2939..2961)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3088..3543
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3175..3194)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3385..3406
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3569..3673
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3674..3742,3743..4534)
                     /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(3892..3911)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4705..5293
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5194..5213
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5581..5602
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5617..5622,5623..5640,5641..5647)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5655..5671
                     /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     5660..5682
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5679..5695
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5679..5695
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 ctgcgcgctc gctcgctcac tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt
       61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact
      121 aggggttcct tgtagttaat gattaacccg ccatgctact tatctaccag ggtaatgggg
      181 atcctctaga actatagcta gccagatata cgcgttgaca ttgattattg actagttatt
      241 aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat
      301 aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa
      361 taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg
      421 agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc
      481 cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct
      541 tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga
      601 tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa
      661 gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc
      721 caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg
      781 aggtctatat aagcagagct cctcgagaag cggccgcact actccgcatg tacccatatg
      841 atgtgcccga ctatgctgaa gacgccaaaa acataaagaa aggcccggcg ccattctatc
      901 cgctggaaga tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg
      961 ttcctggaac aattgctttt acagatgcac atatcgaggt ggacatcact tacgctgagt
     1021 acttcgaaat gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc
     1081 acagaatcgt cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt
     1141 tatttatcgg agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca
     1201 acagtatggg catttcgcag cctaccgtgg tgttcgtttc caaaaagggg ttgcaaaaaa
     1261 ttttgaacgt gcaaaaaaag ctcccaatca tccaaaaaat tattatcatg gattctaaaa
     1321 cggattacca gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt
     1381 ttaatgaata cgattttgtg ccagagtcct tcgataggga caagacaatt gcactgatca
     1441 tgaactcctc tggatctact ggtctgccta aaggtgtcgc tctgcctcat agaactgcct
     1501 gcgtgagatt ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg
     1561 cgattttaag tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt
     1621 tgatatgtgg atttcgagtc gtcttaatgt atagatttga agaagagctg tttctgagga
     1681 gccttcagga ttacaagatt caaagtgcgc tgctggtgcc aaccctattc tccttcttcg
     1741 ccaaaagcac tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctggtg
     1801 gcgctcccct ctctaaggaa gtcggggaag cggttgccaa gaggttccat ctgccaggta
     1861 tcaggcaagg atatgggctc actgagacta catcagctat tctgattaca cccgaggggg
     1921 atgataaacc gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc
     1981 tggataccgg gaaaacgctg ggcgttaatc aaagaggcga actgtgtgtg agaggtccta
     2041 tgattatgtc cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg
     2101 gatggctaca ttctggagac atagcttact gggacgaaga cgaacacttc ttcatcgttg
     2161 accgcctgaa gtctctgatt aagtacaaag gctatcaggt ggctcccgct gaattggaat
     2221 ccatcttgct ccaacacccc aacatcttcg acgcaggtgt cgcaggtctt cccgacgatg
     2281 acgccggtga acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa
     2341 aagagatcgt ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag
     2401 ttgtgtttgt ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca
     2461 gagagatcct cataaaggcc aagaagggcg gaaagatcgc cgtgtaagta cctctagagt
     2521 cgacccgggc ggcctcgagg acggggtgaa ctacgcctga ggatccgatc tttttccctc
     2581 tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg gctaataaag
     2641 gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac tcggaagcaa
     2701 ttcgttgatc tgaatttcga ccacccataa tacccattac cctggtagat aagtagcatg
     2761 gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact ccctctctgc
     2821 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     2881 gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca ctggccgtcg
     2941 ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc cttgcagcac
     3001 atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc ccttcccaac
     3061 agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta agcgcggcgg
     3121 gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg cccgctcctt
     3181 tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa gctctaaatc
     3241 gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc aaaaaacttg
     3301 attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt cgccctttga
     3361 cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca acactcaacc
     3421 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     3481 aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta acgcttacaa
     3541 tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3601 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3661 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3721 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3781 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3841 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3901 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     3961 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     4021 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     4081 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     4141 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4201 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4261 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4321 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4381 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4441 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4501 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4561 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4621 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4681 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4741 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4801 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     4861 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4921 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4981 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5041 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     5101 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     5161 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5221 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5281 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5341 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5401 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5461 cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5521 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5581 taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc ttccggctcg
     5641 tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct atgaccatga
     5701 ttacgccaga tttaattaag gccttaatta gg
//
