LOCUS Exported 6541 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION AAV expression of TurboRFP from CB7 promoter. ACCESSION . VERSION . KEYWORDS pENN.AAV.CB7.CI.TurboRFP.WPRE.RBG SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6541) TITLE AAV plasmids REFERENCE 2 (bases 1 to 6541) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6541 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR enhancer 204..583 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 585..862 /label=chicken beta-actin promoter intron 863..1879 /label=chimeric intron /note="chimera between introns from chicken beta-actin and rabbit beta-globin" primer_bind 1803..1825 /label=pCAGGS-5 /note="Chimeric intron in CAG promoter, forward primer" primer_bind 1887..1906 /label=pCAG-F /note="Rabbit beta-globin intron, for pCAG plasmids, forward primer" CDS 1988..2683 /codon_start=1 /product="red fluorescent protein from Entacmaea quadricolor" /label=TurboRFP /note="mammalian codon-optimized" /translation="MSELIKENMHMKLYMEGTVNNHHFKCTSEGEGKPYEGTQTMKIKV VEGGPLPFAFDILATSFMYGSKAFINHTQGIPDFFKQSFPEGFTWERITTYEDGGVLTA TQDTSFQNGCIIYNVKINGVNFPSNGPVMQKKTRGWEANTEMLYPADGGLRGHSQMALK LVGGGYLHCSFKTTYRSKKPAKNLKMPGFHFVDHRLERIKEADKETYVEQHEMAVAKYC DLPSKLGHR" misc_feature 2743..3331 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(2796..2816) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3214..3225) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3378..3397) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3443..3498 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" repeat_region 3591..3720 /label=AAV2 ITR primer_bind complement(3739..3756) /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind complement(3739..3755) /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind complement(3748..3770) /label=M13/pUC Forward /note="In lacZ gene" rep_origin 3897..4352 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(3984..4003) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4194..4215 /label=F1ori-F /note="F1 origin, forward primer" promoter 4378..4482 /gene="bla" /label=AmpR promoter CDS 4483..5343 /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(4701..4720) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 5514..6102 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6003..6022 /label=pBR322ori-F /note="pBR322 origin, forward primer" primer_bind 6256..6273 /label=L4440 /note="L4440 vector, forward primer" protein_bind 6390..6411 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." promoter 6426..6456 /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 6464..6480 /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 6469..6491 /label=M13/pUC Reverse /note="In lacZ gene" primer_bind 6488..6504 /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind 6488..6504 /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 gtcgacattg attattgact agttattaat agtaatcaat 241 tacggggtca ttagttcata gcccatatat ggagttccgc gttacataac ttacggtaaa 301 tggcccgcct ggctgaccgc ccaacgaccc ccgcccattg acgtcaataa tgacgtatgt 361 tcccatagta acgccaatag ggactttcca ttgacgtcaa tgggtggagt atttacggta 421 aactgcccac ttggcagtac atcaagtgta tcatatgcca agtacgcccc ctattgacgt 481 caatgacggt aaatggcccg cctggcatta tgcccagtac atgaccttat gggactttcc 541 tacttggcag tacatctacg tattagtcat cgctattacc atggtcgagg tgagccccac 601 gttctgcttc actctcccca tctccccccc ctccccaccc ccaattttgt atttatttat 661 tttttaatta ttttgtgcag cgatgggggc gggggggggg ggggggcgcg cgccaggcgg 721 ggcggggcgg ggcgaggggc ggggcggggc gaggcggaga ggtgcggcgg cagccaatca 781 gagcggcgcg ctccgaaagt ttccttttat ggcgaggcgg cggcggcggc ggccctataa 841 aaagcgaagc gcgcggcggg cgggagtcgc tgcgcgctgc cttcgccccg tgccccgctc 901 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag 961 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt 1021 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg 1081 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc 1141 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt 1201 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa 1261 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt 1321 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg 1381 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca 1441 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg 1501 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt 1561 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa 1621 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg 1681 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc 1741 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc 1801 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc 1861 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt 1921 tggcaaagaa tagcttcgaa ttctgcagtc gacggtaccg cgggcccggg atccaccggt 1981 cgccaccatg agcgagctga tcaaggagaa catgcacatg aagctgtaca tggagggcac 2041 cgtgaacaac caccacttca agtgcacatc cgagggcgaa ggcaagccct acgagggcac 2101 ccagaccatg aagatcaagg tggtcgaggg cggccctctc cccttcgcct tcgacatcct 2161 ggctaccagc ttcatgtacg gcagcaaagc cttcatcaac cacacccagg gcatccccga 2221 cttctttaag cagtccttcc ctgagggctt cacatgggag agaatcacca catacgaaga 2281 cgggggcgtg ctgaccgcta cccaggacac cagcttccag aacggctgca tcatctacaa 2341 cgtcaagatc aacggggtga acttcccatc caacggccct gtgatgcaga agaaaacacg 2401 cggctgggag gccaacaccg agatgctgta ccccgctgac ggcggcctga gaggccacag 2461 ccagatggcc ctgaagctcg tgggcggggg ctacctgcac tgctccttca agaccacata 2521 cagatccaag aaacccgcta agaacctcaa gatgcccggc ttccacttcg tggaccacag 2581 actggaaaga atcaaggagg ccgacaaaga gacctacgtc gagcagcacg agatggctgt 2641 ggccaagtac tgcgacctcc ctagcaaact ggggcacaga tgatgcggcc cgcgtggtac 2701 ctctagagtc gacccgggcg gcctcgaatc aagcttatcg ataatcaacc tctggattac 2761 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga 2821 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc 2881 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa 2941 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc 3001 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc 3061 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc 3121 gtggtgttgt cggggaaatc atcgtccttt ccttggctgc tcgcctgtgt tgccacctgg 3181 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct 3241 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg 3301 agtcggatct ccctttgggc cgcctccccg catcgatacc gtctcgagga cggggtgaac 3361 tacgcctgag gatccgatct ttttccctct gccaaaaatt atggggacat catgaagccc 3421 cttgagcatc tgacttctgg ctaataaagg aaatttattt tcattgcaat agtgtgttgg 3481 aattttttgt gtctctcact cggaagcaat tcgttgatct gaatttcgac cacccataat 3541 acccattacc ctggtagata agtagcatgg cgggttaatc attaactaca aggaacccct 3601 agtgatggag ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc 3661 aaaggtcgcc cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag 3721 ccttaattaa cctaattcac tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg 3781 cgttacccaa cttaatcgcc ttgcagcaca tccccctttc gccagctggc gtaatagcga 3841 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc 3901 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac 3961 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt 4021 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc 4081 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc 4141 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact 4201 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg 4261 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc 4321 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc 4381 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa 4441 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc 4501 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa 4561 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa 4621 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg 4681 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa 4741 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc 4801 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc 4861 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta 4921 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag 4981 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca 5041 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata 5101 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc 5161 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca 5221 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca 5281 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg 5341 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa 5401 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt 5461 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat 5521 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg 5581 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga 5641 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac 5701 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt 5761 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag 5821 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc 5881 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag 5941 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca 6001 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt 6061 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc 6121 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc 6181 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc 6241 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa 6301 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac 6361 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc 6421 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa 6481 tttcacacag gaaacagcta tgaccatgat tacgccagat ttaattaagg ccttaattag 6541 g //