LOCUS       Exported                5322 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  AAV expression of TurboRFP from CMV promoter.
ACCESSION   .
VERSION     .
KEYWORDS    pENN.AAV.CMVs.TurboRFP.WPRE.RBG
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5322)
  TITLE     AAV plasmids
REFERENCE   2  (bases 1 to 5322)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5322
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..130
                     /label=AAV2 ITR
     promoter        485..688
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     639..659
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     685..709
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     CDS             808..1503
                     /codon_start=1
                     /product="red fluorescent protein from Entacmaea 
                     quadricolor"
                     /label=TurboRFP
                     /note="mammalian codon-optimized"
                     /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV
                     VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA
                     TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK
                     LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC
                     DLPSKLGHR"
     misc_feature    1524..2112
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(1577..1597)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     CDS             complement(1995..2006)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(2159..2178)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2224..2279
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     repeat_region   2372..2501
                     /label=AAV2 ITR
     primer_bind     complement(2520..2537)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(2520..2536)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2529..2551)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2678..3133
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2765..2784)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2975..2996
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3159..3263
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3264..4124
                     /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(3482..3501)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4295..4883
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4784..4803
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5037..5054
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5171..5192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        5207..5237
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5245..5261
                     /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     5250..5272
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5269..5285
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5269..5285
                     /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 gcatgcctgc aggctgaccg cccaacgacc cccgcccatt
      241 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      301 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      361 aagtccgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      421 catgacctta cgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      481 catggtgatg cggttttggc agtacaccaa tgggcgtgga tagcggtttg actcacgggg
      541 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      601 ggactttcca aaatgtcgta ataaccccgc cccgttgacg caaatgggcg gtaggcgtgt
      661 acggtgggag gtctatataa gcagagctcg tttagtgaac cgtcagatca ctagaagctt
      721 tattgcggta gtttatcaca gttaaattgc taacgcagtc agtgcttctg acacaacagt
      781 ctcgaactta atcgaactag tgccaccatg agcgagctga tcaaggagaa catgcacatg
      841 aagctgtaca tggagggcac cgtgaacaac caccacttca agtgcacatc cgagggcgaa
      901 ggcaagccct acgagggcac ccagaccatg aagatcaagg tggtcgaggg cggccctctc
      961 cccttcgcct tcgacatcct ggctaccagc ttcatgtacg gcagcaaagc cttcatcaac
     1021 cacacccagg gcatccccga cttctttaag cagtccttcc ctgagggctt cacatgggag
     1081 agaatcacca catacgaaga cgggggcgtg ctgaccgcta cccaggacac cagcttccag
     1141 aacggctgca tcatctacaa cgtcaagatc aacggggtga acttcccatc caacggccct
     1201 gtgatgcaga agaaaacacg cggctgggag gccaacaccg agatgctgta ccccgctgac
     1261 ggcggcctga gaggccacag ccagatggcc ctgaagctcg tgggcggggg ctacctgcac
     1321 tgctccttca agaccacata cagatccaag aaacccgcta agaacctcaa gatgcccggc
     1381 ttccacttcg tggaccacag actggaaaga atcaaggagg ccgacaaaga gacctacgtc
     1441 gagcagcacg agatggctgt ggccaagtac tgcgacctcc ctagcaaact ggggcacaga
     1501 tgactcgaat caagcttatc gataatcaac ctctggatta caaaatttgt gaaagattga
     1561 ctggtattct taactatgtt gctcctttta cgctatgtgg atacgctgct ttaatgcctt
     1621 tgtatcatgc tattgcttcc cgtatggctt tcattttctc ctccttgtat aaatcctggt
     1681 tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca acgtggcgtg gtgtgcactg
     1741 tgtttgctga cgcaaccccc actggttggg gcattgccac cacctgtcag ctcctttccg
     1801 ggactttcgc tttccccctc cctattgcca cggcggaact catcgccgcc tgccttgccc
     1861 gctgctggac aggggctcgg ctgttgggca ctgacaattc cgtggtgttg tcggggaaat
     1921 catcgtcctt tccttggctg ctcgcctgtg ttgccacctg gattctgcgc gggacgtcct
     1981 tctgctacgt cccttcggcc ctcaatccag cggaccttcc ttcccgcggc ctgctgccgg
     2041 ctctgcggcc tcttccgcgt cttcgccttc gccctcagac gagtcggatc tccctttggg
     2101 ccgcctcccc gcatcgatac cgtctcgagg acggggtgaa ctacgcctga ggatccgatc
     2161 tttttccctc tgccaaaaat tatggggaca tcatgaagcc ccttgagcat ctgacttctg
     2221 gctaataaag gaaatttatt ttcattgcaa tagtgtgttg gaattttttg tgtctctcac
     2281 tcggaagcaa ttcgttgatc tgaatttcga ccacccataa tacccattac cctggtagat
     2341 aagtagcatg gcgggttaat cattaactac aaggaacccc tagtgatgga gttggccact
     2401 ccctctctgc gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg
     2461 ggctttgccc gggcggcctc agtgagcgag cgagcgcgca gccttaatta acctaattca
     2521 ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg gcgttaccca acttaatcgc
     2581 cttgcagcac atcccccttt cgccagctgg cgtaatagcg aagaggcccg caccgatcgc
     2641 ccttcccaac agttgcgcag cctgaatggc gaatgggacg cgccctgtag cggcgcatta
     2701 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     2761 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     2821 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     2881 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     2941 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3001 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3061 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa caaaatatta
     3121 acgcttacaa tttaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3181 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3241 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3301 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3361 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3421 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3481 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3541 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3601 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3661 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3721 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3781 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3841 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3901 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     3961 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4021 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4081 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4141 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4201 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4261 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4321 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4381 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4441 ttcttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4501 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4561 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4621 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4681 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4741 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4801 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4861 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4921 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     4981 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5041 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5101 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5161 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5221 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5281 atgaccatga ttacgccaga tttaattaag gccttaatta gg
//
