LOCUS       sequence-202402-        5300 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  AAV expression of TurboRFP from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5300)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5300)
  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.CMVs.TurboRFP.WPRE.RBG
FEATURES             Location/Qualifiers
     source          1..5300
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        471..674
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     625..645
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     671..695
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             794..1489
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     misc_feature    1510..2098
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1563..1583)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1981..1992)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2145..2164)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2210..2265
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2358..2487
                     /label=AAV2 ITR
     primer_bind     complement(2506..2523)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2506..2522)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2515..2537)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2664..3119
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2751..2770)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2961..2982
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3145..3249
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3250..3318,3319..4110)
                     /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(3468..3487)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4281..4869
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4770..4789
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     protein_bind    5157..5178
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5193..5198,5199..5216,5217..5223)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5231..5247
                     /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     5236..5258
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5255..5271
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5255..5271
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 taattaggct gcgcgctcgc tcgctcactg aggccgcccg ggcaaagccc gggcggcctc
       61 agtgagcgag cgagcgcgca gagagggagt ggccaactcc atcactaggg gttccttgta
      121 gttaatgatt aacccgccat gctacttatc taccagggta atggggatcc tctagaacta
      181 tagctagcat gcctgcaggc tgaccgccca acgacccccg cccattgacg tcaataatga
      241 cgtatgttcc catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt
      301 tacggtaaac tgcccacttg gcagtacatc aagtgtatca tatgccaagt ccgcccccta
      361 ttgacgtcaa tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttacggg
      421 actttcctac ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt
      481 tttggcagta caccaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc
      541 accccattga cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat
      601 gtcgtaataa ccccgccccg ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct
      661 atataagcag agctcgttta gtgaaccgtc agatcactag aagctttatt gcggtagttt
      721 atcacagtta aattgctaac gcagtcagtg cttctgacac aacagtctcg aacttaatcg
      781 aactagtgcc accatgagcg agctgatcaa ggagaacatg cacatgaagc tgtacatgga
      841 gggcaccgtg aacaaccacc acttcaagtg cacatccgag ggcgaaggca agccctacga
      901 gggcacccag accatgaaga tcaaggtggt cgagggcggc cctctcccct tcgccttcga
      961 catcctggct accagcttca tgtacggcag caaagccttc atcaaccaca cccagggcat
     1021 ccccgacttc tttaagcagt ccttccctga gggcttcaca tgggagagaa tcaccacata
     1081 cgaagacggg ggcgtgctga ccgctaccca ggacaccagc ttccagaacg gctgcatcat
     1141 ctacaacgtc aagatcaacg gggtgaactt cccatccaac ggccctgtga tgcagaagaa
     1201 aacacgcggc tgggaggcca acaccgagat gctgtacccc gctgacggcg gcctgagagg
     1261 ccacagccag atggccctga agctcgtggg cgggggctac ctgcactgct ccttcaagac
     1321 cacatacaga tccaagaaac ccgctaagaa cctcaagatg cccggcttcc acttcgtgga
     1381 ccacagactg gaaagaatca aggaggccga caaagagacc tacgtcgagc agcacgagat
     1441 ggctgtggcc aagtactgcg acctccctag caaactgggg cacagatgac tcgaatcaag
     1501 cttatcgata atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     1561 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     1621 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     1681 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     1741 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     1801 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     1861 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaaatcatc gtcctttcct
     1921 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     1981 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     2041 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcat
     2101 cgataccgtc tcgaggacgg ggtgaactac gcctgaggat ccgatctttt tccctctgcc
     2161 aaaaattatg gggacatcat gaagcccctt gagcatctga cttctggcta ataaaggaaa
     2221 tttattttca ttgcaatagt gtgttggaat tttttgtgtc tctcactcgg aagcaattcg
     2281 ttgatctgaa tttcgaccac ccataatacc cattaccctg gtagataagt agcatggcgg
     2341 gttaatcatt aactacaagg aacccctagt gatggagttg gccactccct ctctgcgcgc
     2401 tcgctcgctc actgaggccg ggcgaccaaa ggtcgcccga cgcccgggct ttgcccgggc
     2461 ggcctcagtg agcgagcgag cgcgcagcct taattaacct aattcactgg ccgtcgtttt
     2521 acaacgtcgt gactgggaaa accctggcgt tacccaactt aatcgccttg cagcacatcc
     2581 ccctttcgcc agctggcgta atagcgaaga ggcccgcacc gatcgccctt cccaacagtt
     2641 gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     2701 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     2761 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     2821 gctcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta
     2881 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     2941 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     3001 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     3061 tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa atattaacgc ttacaattta
     3121 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     3181 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     3241 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     3301 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     3361 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     3421 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     3481 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     3541 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     3601 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     3661 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     3721 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     3781 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     3841 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     3901 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     3961 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     4021 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     4081 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     4141 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     4201 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     4261 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     4321 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     4381 tttccgaagg taactggctt cagcagagcg cagataccaa atactgttct tctagtgtag
     4441 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     4501 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     4561 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     4621 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gctatgagaa
     4681 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     4741 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     4801 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     4861 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     4921 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     4981 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     5041 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     5101 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     5161 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     5221 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     5281 gccagattta attaaggcct
//
