LOCUS Exported 6727 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Fluorescent Reporter for Dopaminergic Neuron Differentiation. ACCESSION . VERSION . KEYWORDS pAAV2.5-THP-GFP SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6727) AUTHORS Oh MS, Hong SJ, Huh Y, Kim KS TITLE Expression of transgenes in midbrain dopamine neurons using the tyrosine hydroxylase promoter. JOURNAL Gene Ther. 2009 Mar;16(3):437-40. doi: 10.1038/gt.2008.148. Epub 2008 Sep 18. PUBMED 18800154 REFERENCE 2 (bases 1 to 6727) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6727 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..130 /label=AAV2 ITR (alternate) /note="Functional equivalent of wild-type AAV2 ITR" regulatory 2694..2703 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 2700..3419 /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 2700..3416 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2745..2766) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3006..3025) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3353..3374 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 3474..3950 /label=hGH poly(A) signal /note="human growth hormone polyadenylation signal" primer_bind complement(3673..3692) /label=hGH-PA-R /note="Human growth hormone terminator, reverse primer" repeat_region 3990..4130 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 3990..4119 /label=AAV2 ITR rep_origin 4205..4660 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4292..4311) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4502..4523 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4677..4696) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4796..4818 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4856..4874) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4942..5046 /gene="bla" /label=AmpR promoter CDS 5047..5907 /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(5265..5284) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6078..6666 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6567..6586 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcgtcg ggcgaccttt 61 ggtcgcccgg cctcagtgag cgagcgagcg cgcagagagg gagtggccaa ctccatcact 121 aggggttcct gcggccgcac gcgaggcctc ttgggattct tctcatcaaa aagggcatgg 181 gacacagcta aagcgtccag ggctcctctg tgcccacaga tgccttagat cttggcacaa 241 tgtagtcagc cagctccgtg tgtgtgtgtg tgtgtttgca tgtatctcac agacagtgca 301 caatggcctg gatgtgaaca gaggcaagag tctgggccag cagttgtctc ccaggagggt 361 ccaaagacat cgtattttca agtttaggcc aggtgctcac ttgggtgagc tcagacacag 421 acaaaggtct ggagagcaca cattccccac ccccacccag ctcctatgca agcacctcca 481 gccgagacaa gaaaacgaat taaaaagcaa tatttgtgtc agcgtaagac atttgccgaa 541 aggttaaatc cacactcgtg gtgctgcaca gcagccccct gtgcaggatt tgttaggcac 601 agctccctcc taccccgtgc cacctgagca aatgccaggc tgggtgggct ggaaccagcc 661 tgggcttgcc tcaccctgga atccccagca ccctccaaag gaggaccctg ggagtgggca 721 tagacgccct tcaggtgtgg gcaacagccc ccagtcctca ggatgaaagg ctaaggtgca 781 gccagctctg ccttcacggt gggaatgtct ctatgtgagc cctttctggg ctgtgaagaa 841 cgctctgaga agggtcctgg gaccctggat aggccagagc tgtgctgggc atgtagagac 901 aggagtgggc taaagcagca aaggcactga ccaaggaaga gttcagagag gagcgtggaa 961 tatggggagg ggttcatagt aagagagagc aggcagtgga gagtaaatag tcactgagcc 1021 ggggtttatg gggtttgtag gagcttactc agagaaagta gatgagagat gccatgccag 1081 tctgagtatc acagagcccc aggctctcct gggaacggaa ctgtgagggc cagaaggtca 1141 gcaagggagg ttagggagag ttccttttgt actgactcag catttatcct gctcccaggg 1201 ggcaatgggg gccagtgagg gatgcagagc aaggcagtga tgtggcaggc agttcctgtt 1261 gtgaaagagc tgggaaggga gcgggctggg cctggtacgt acagcaggcc atttctgagg 1321 gtccgagtgc tgtctaggag gtgcagtgag acttcagtga tcagccagaa cagaagctaa 1381 gcggggtggg gactgcgagt tcaggcttct gggtcttgca aatatccaga atgctaaatc 1441 ctcagaaccc cagggtggcc attttcagag tgggttttgt cctttgggca cttgtgcaga 1501 ctccaatatc cagagggata aggatggtac tcttcagtac ccttagtgag aggacacttt 1561 tctctgaagg gcttgaatgt gccgagccat tacctgaagg aaggaaatga ctccagggac 1621 ataggatggg cccagcacaa ctcacctgct acagagaaag gtcccctccc tggtctcctt 1681 agagatcctg tttccctggc tgaggaagct agggtggatc tttgtgtaag tgggtgtgga 1741 tgctaactgg aaaacaaaag gtcacttact gttagacctc ggggtaccat ggaagagatg 1801 atcactgagt gtgcccttac atggggacca gctgagaatg gggctaccac tagctcgaga 1861 ccatgataca gggaataagt gtgcatttgg gggtagggag tggctcagaa tactcttaac 1921 caaagcagag gtttgctccc acaggaaggt gaggtcagaa ggccttaggg agctgccagg 1981 ggctagggtt ggcaccatct cccaggctgt gtctttaagg agatgataat cagagggata 2041 gaaccttgca aaagtgggcc agtcttggga atactataga ggaatagcct tctggaacat 2101 tctgtgtctc ataggacctg cctggggatc cagccccagt gccagcacat ataccgactg 2161 gggcagtgaa tagatagtac actttgttac atgggctggg gggaacatgg cccatgtcct 2221 ggaggggact ttatgacaga catccaaaaa tccagtgaga gggcttctag atttgtctcc 2281 aaaggttata gttctaacat gagcccttag gaaatccagc atggttctcc ctgtgtgccc 2341 tggtttggtt agagagctct agcggtctcc tgtcccacag aataccagcc agcccctgcc 2401 ctacgtcgtg cctcgggctg agggtgattc agaggcaggt gcctgtgaca gtggatgcaa 2461 ttagatctaa tgggacggag gcctttctcg tcgccctcgc tccatgccca cccccgcctc 2521 cctcaggcac agcaggcgtg gagaggatgc gcaggaggta ggaggtgggg gacccagagg 2581 ggctttgacg tcagcctggc ctttaaagag ggcgcctgcc tggcgagggc tgtggagaca 2641 gaactcggga ccaccagagg aattcgagct cggtacccgg ggatcctcta gacgccacca 2701 tggtgagcaa gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg 2761 gcgacgtaaa cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg 2821 gcaagctgac cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc 2881 tcgtgaccac cctgacctac ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc 2941 agcacgactt cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct 3001 tcaaggacga cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg 3061 tgaaccgcat cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca 3121 agctggagta caactacaac agccacaacg tctatatcat ggccgacaag cagaagaacg 3181 gcatcaaggt gaacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg 3241 accactacca gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact 3301 acctgagcac ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc 3361 tgctggagtt cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtaat 3421 ctagagtcga cctgcagaag cttgcctcga gcagcgctgc tcgagagatc tacgggtggc 3481 atccctgtga cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca 3541 ccagccttgt cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata 3601 atattatggg gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta 3661 gggcctgcgg ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg 3721 caatctccgc ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat 3781 tccaggcatg catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca 3841 ccatattggc caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc 3901 ccaaattgct gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttgt 3961 aggtaaccac gtgcggaccg agcggccgca ggaaccccta gtgatggagt tggccactcc 4021 ctctctgcgc gctcgctcgc tcactgaggc cgggcgacca aaggtcgccc gacgcccggg 4081 ctttgcccgg gcggcctcag tgagcgagcg agcgcgcagc tgcctgcagg ggcgcctgat 4141 gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatacgtc aaagcaacca 4201 tagtacgcgc cctgtagcgg cgcattaagc gcggcgggtg tggtggttac gcgcagcgtg 4261 accgctacac ttgccagcgc cctagcgccc gctcctttcg ctttcttccc ttcctttctc 4321 gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg ggctcccttt agggttccga 4381 tttagtgctt tacggcacct cgaccccaaa aaacttgatt tgggtgatgg ttcacgtagt 4441 gggccatcgc cctgatagac ggtttttcgc cctttgacgt tggagtccac gttctttaat 4501 agtggactct tgttccaaac tggaacaaca ctcaacccta tctcgggcta ttcttttgat 4561 ttataaggga ttttgccgat ttcggcctat tggttaaaaa atgagctgat ttaacaaaaa 4621 tttaacgcga attttaacaa aatattaacg tttacaattt tatggtgcac tctcagtaca 4681 atctgctctg atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg 4741 ccctgacggg cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg 4801 agctgcatgt gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc 4861 gtgatacgcc tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt 4921 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca 4981 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg 5041 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc 5101 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg 5161 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt 5221 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta 5281 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat 5341 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga 5401 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca 5461 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact 5521 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc 5581 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact 5641 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt 5701 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt 5761 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt 5821 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata 5881 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag 5941 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat 6001 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa 6061 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca 6121 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt 6181 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct agtgtagccg 6241 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc 6301 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga 6361 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc 6421 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc 6481 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca 6541 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg 6601 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta 6661 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct 6721 cacatgt //