LOCUS       sequence-202437-        5573 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of EGFP from cTNT promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5573)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5573)
  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"
COMMENT     Sequence Label: pENN.AAV.cTNT.PI.eGFP.WPRE.rBG
FEATURES             Location/Qualifiers
     source          1..5573
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    113..134
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(149..154,155..172,173..179)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    187..203
                     /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     192..214
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     211..227
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     211..227
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     repeat_region   265..394
                     /label=AAV2 ITR
     intron          1036..1168
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     primer_bind     1151..1170
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        1213..1231
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      1254..1263
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(1260..1262,1263..1265,1266..1979)
                     /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(1260..1262,1263..1265,1266..1976)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1305..1326)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1566..1585)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1913..1934
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     misc_feature    2060..2648
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(2113..2133)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(2531..2542)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2674..2693)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2739..2794
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2887..3016
                     /label=AAV2 ITR
     primer_bind     complement(3035..3052)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3035..3051)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3044..3066)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      3193..3648
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3280..3299)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3490..3511
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3674..3778
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3779..3847,3848..4639)
                     /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(3997..4016)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4810..5398
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5299..5318
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
       61 gctggcacga caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga
      121 gttagctcac tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt
      181 gtggaattgt gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca
      241 gatttaatta aggccttaat taggctgcgc gctcgctcgc tcactgaggc cgcccgggca
      301 aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag tgagcgagcg agcgcgcaga
      361 gagggagtgg ccaactccat cactaggggt tccttgtagt taatgattaa cccgccatgc
      421 tacttatcta ccagggtaat ggggatcctc tagaactata gctagaattc gcccttacgg
      481 gccccccctc gaggtcggga taaaagcagt ctgggctttc acatgacagc atctggggct
      541 gcggcagagg gtcgggtccg aagcgctgcc ttatcagcgt ccccagccct gggaggtgac
      601 agctggctgg cttgtgtcag cccctcgggc actcacgtat ctccgtccga cgggtttaaa
      661 atagcaaaac tctgaggcca cacaatagct tgggcttata tgggctcctg tgggggaagg
      721 gggagcacgg agggggccgg ggccgctgct gccaaaatag cagctcacaa gtgttgcatt
      781 cctctctggg cgccgggcac attcctgctg gctctgcccg ccccggggtg ggcgccgggg
      841 ggaccttaaa gcctctgccc cccaaggagc ccttcccaga cagccgccgg cacccaccgc
      901 tccgtgggac gatccccgaa gctctagagc tttattgcgg tagtttatca cagttaaatt
      961 gctaacgcag tcagtgcttc tgacacaaca gtctcgaact taagctgcag aagttggtcg
     1021 tgaggcactg ggcaggtaag tatcaaggtt acaagacagg tttaaggaga ccaatagaaa
     1081 ctgggcttgt cgagacagag aagactcttg cgtttctgat aggcacctat tggtcttact
     1141 gacatccact ttgcctttct ctccacaggt gtccactccc agttcaatta cagctcttaa
     1201 ggctagagta cttaatacga ctcactatag gctagcctcg agaattcacg cgggccgcca
     1261 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg
     1321 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg
     1381 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc
     1441 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc
     1501 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct
     1561 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg
     1621 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca
     1681 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg
     1741 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg
     1801 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact
     1861 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc
     1921 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat
     1981 aagctcgcgt ggtacctcta gagtcgaccc gggcggcctc gaggacgggg tgaactacgc
     2041 ctgaatcaag cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg
     2101 tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta
     2161 tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct
     2221 gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt
     2281 tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac
     2341 tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg
     2401 ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc
     2461 gtcctttcct tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg
     2521 ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct
     2581 gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc
     2641 ctccccgcat cgataccgtc gatgaggatc cgatcttttt ccctctgcca aaaattatgg
     2701 ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat
     2761 tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga agcaattcgt tgatctgaat
     2821 ttcgaccacc cataataccc attaccctgg tagataagta gcatggcggg ttaatcatta
     2881 actacaagga acccctagtg atggagttgg ccactccctc tctgcgcgct cgctcgctca
     2941 ctgaggccgg gcgaccaaag gtcgcccgac gcccgggctt tgcccgggcg gcctcagtga
     3001 gcgagcgagc gcgcagcctt aattaaccta attcactggc cgtcgtttta caacgtcgtg
     3061 actgggaaaa ccctggcgtt acccaactta atcgccttgc agcacatccc cctttcgcca
     3121 gctggcgtaa tagcgaagag gcccgcaccg atcgcccttc ccaacagttg cgcagcctga
     3181 atggcgaatg ggacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc
     3241 gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct ttcttccctt
     3301 cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg ctccctttag
     3361 ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag ggtgatggtt
     3421 cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg gagtccacgt
     3481 tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc tcggtctatt
     3541 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     3601 aacaaaaatt taacgcgaat tttaacaaaa tattaacgct tacaatttag gtggcacttt
     3661 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3721 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3781 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     3841 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     3901 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     3961 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4021 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4081 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4141 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4201 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4261 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4321 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4381 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4441 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4501 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4561 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4621 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4681 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4741 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4801 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     4861 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     4921 aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc cgtagttagg
     4981 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5041 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5101 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5161 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5221 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5281 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5341 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5401 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     5461 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     5521 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aag
//
