LOCUS       sequence-485255-        4593 bp    DNA     circular SYN 03-SEP-2025
DEFINITION  AAV plasmid expressing HA-EGFP, control for plasmid 242960.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4593)
  TITLE     pAG mouse IgG2a and AAV Plasmids
  JOURNAL   Unpublished
REFERENCE   2  (bases 1 to 4593)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Sep 3, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: 
            "Unpublished"
COMMENT     Sequence Label: pAG AAV CMV HA-EGFP
FEATURES             Location/Qualifiers
     source          1..4593
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     100..122
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     114..131
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     115..131
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     141..160
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        141..159
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     183..199
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     184..200
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     repeat_region   210..350
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   221..350
                     /label=AAV2 ITR
     promoter        442..644
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     596..616
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     CDS             699..725
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(732..734,735..737,738..1448)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             join(732..734,735..737,738..1448)
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(777..798)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1038..1057)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1385..1406
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    1541..1662
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1578..1597)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1632..1651
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     repeat_region   1689..1829
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   1689..1818
                     /label=AAV2 ITR
     primer_bind     complement(1920..1940)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1920..1938)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1959..1975)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1959..1975)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1972..1994)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1983..1999
                     /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)."
     promoter        complement(join(2007..2013,2014..2031,2032..2037))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2052..2073
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     rep_origin      complement(2361..2949)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2441..2460)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3120..3911,3912..3980))
                     /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     3743..3762
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3981..4085)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(4111..4566)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4248..4269)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     4460..4479
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
ORIGIN
        1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat
       61 gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga cgttgtaaaa
      121 cgacggccag tgagcgcgcg taatacgact cactataggg cgaatgggta ccggaccccc
      181 cctcgaggtc gacggtatcg ggggcatgcc ctgcaggcag ctgcgcgctc gctcgctcac
      241 tgaggccgcc cgggcaaagc ccgggcgtcg ggcgaccttt ggtcgcccgg cctcagtgag
      301 cgagcgagcg cgcagagagg gagtggccaa ctccatcact aggggttcct gcggccgcag
      361 ccgtgtttta gagctagaaa tagcaagtta aaataaggct agtccgtagc gcgtgcgcca
      421 attctgcaga caaattctag agtgatgcgg ttttggcagt acatcaatgg gcgtggatag
      481 cggtttgact cacggggatt tccaagtctc caccccattg acgtcaatgg gagtttgttt
      541 tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa
      601 atgggcggta ggcgtgtacg gtgggaggtc tatataagca gagcggtcac cggtttagtg
      661 aaccgtcaga tccgctagcg ctaccggtcg ccaccatgta cccatacgat gttccggatt
      721 atgctttcga aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg
      781 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg
      841 atgccaccta cggcaagctg accctgaagt tcatctgcac caccggcaag ctgcccgtgc
      901 cctggcccac cctcgtgacc accctgacct acggcgtgca gtgcttcagc cgctaccccg
      961 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc
     1021 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg
     1081 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca
     1141 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc atggccgaca
     1201 agcagaagaa cggcatcaag gtgaacttca agatccgcca caacatcgag gacggcagcg
     1261 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc
     1321 ccgacaacca ctacctgagc acccagtccg ccctgagcaa agaccccaac gagaagcgcg
     1381 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc
     1441 tgtacaaggg atcctgaacg cgtgtagtag ttcaagtcat cttattattc agtatttata
     1501 acttgcaaag aaatgaatat cagagagtga gaggtgtaca aacttgttta ttgcagctta
     1561 taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat ttttttcact
     1621 gcattctagt tgtggtttgt ccaaactcat caatgtatct tagttaacac gtgcgaccga
     1681 gcggccgcag gaacccctag tgatggagtt ggccactccc tctctgcgcg ctcgctcgct
     1741 cactgaggcc gggcgaccaa aggtcgcccg acgcccgggc tttgcccggg cggcctcagt
     1801 gagcgagcga gcgcgcagct gcctgcaggg gcgccaagcc gaattctgca gatatccatc
     1861 acactggcgg ccgctcgact agagcggccg ccaccgcggt ggagctccag cttttgttcc
     1921 ctttagtgag ggttaattgc gcgcttggcg taatcatggt catagctgtt tcctgtgtga
     1981 aattgttatc cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc
     2041 tggggtgcct aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc
     2101 cagtcgggaa acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc
     2161 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2221 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2281 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2341 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     2401 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     2461 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     2521 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     2581 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     2641 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     2701 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     2761 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     2821 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     2881 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     2941 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3001 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3061 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3121 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3181 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3241 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3301 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3361 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     3421 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     3481 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     3541 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     3601 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     3661 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     3721 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     3781 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     3841 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     3901 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     3961 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4021 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4081 ccgcgcacat ttccccgaaa agtgccacct aaattgtaag cgttaatatt ttgttaaaat
     4141 tcgcgttaaa tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa
     4201 tcccttataa atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca
     4261 agagtccact attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg
     4321 gcgatggccc actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta
     4381 aagcactaaa tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg
     4441 cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa
     4501 gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg
     4561 gcgcgtccca ttcgccattc aggctgcgca act
//
