LOCUS Exported 6131 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION RMCE vector for generating C-terminal T2A-lexA-p65 fusion. ACCESSION . VERSION . KEYWORDS pF3BGX-T2A-lexAp65 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6131) TITLE Neurochemical organization of the Drosophila brain visualized by endogenously tagged neurotransmitter receptors JOURNAL Unpublished REFERENCE 2 (bases 1 to 6131) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6131 /organism="synthetic DNA construct" /mol_type="other DNA" CDS 312..1031 /codon_start=1 /product="yellow fluorescent protein (YFP) with fast and efficient maturation (Nagai et al., 2002)" /label=Venus /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KLICTTGKLPVPWPTLVTTLGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIK ANFKIRHNIEDGGVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(357..378) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(618..637) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 965..986 /label=EGFP-C /note="EGFP, forward primer" polyA_signal 1147..1228 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(1184..1203) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 1238..1257 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" protein_bind 1275..1308 /label=attB /bound_moiety="phage phi-C31 integrase" /note="minimal attB site for the phi-C31 integrase (Groth et al., 2000)" CDS 1320..1373 /codon_start=1 /product="2A peptide from Thosea asigna virus capsid protein" /label=T2A /note="Eukaryotic ribosomes fail to insert a peptide bond between the Gly and Pro residues, yielding separate polypeptides." /translation="EGRGSLLTCGDVEENPGP" CDS 2424..2795 /codon_start=1 /product="transcriptional activation domain of human RelA, also known as p65 (O'Shea and Perkins, 2008)" /label=RelA (p65) AD /translation="PTQAGEGTLSEALLQLQFDDEDLGALLGNSTDPAVFTDLASVDNS EFQQLLNQGIPVAPHTTEPMLMEYPEAITRLVTGAQRPPDPAPAPLGAPGLPNGLLSGD EDFSSIADMDFSALLSQISS" protein_bind 2807..2840 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2913..3145 /label=P element 3' end /note="P element 3' end" primer_bind complement(3251..3268) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(3422..4010) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(3502..3521) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(4181..5041) /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 4804..4823 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(5042..5146) /gene="bla" /label=AmpR promoter primer_bind complement(5309..5330) /label=F1ori-F /note="F1 origin, forward primer" misc_feature 5546..6131 /label=P element 5' end ORIGIN 1 ggtaccgggc cccccctcga ggtcgacgaa gttcctattc ttcaaatagt ataggaactt 61 caagcttctt aaggaattcg agctcgcccg gggatctaat tcaattagag actaattcaa 121 ttagagctaa ttcaattagg atcagcttat cgatttcgaa ccctcgaccg ccggagtata 181 aatagaggcg cttcgtctac ggagcgacaa ttcaattcaa acaagcaaag tgaacacgtc 241 gctaagcgaa agctaagcaa ataaacaagc gcagctgaac aagctaaaca atcgggctag 301 acctaggcaa aatggtgagc aagggcgagg agctgttcac cggggtggtg cccatcctgg 361 tcgagctgga cggcgacgta aacggccaca agttcagcgt gtccggcgag ggcgagggcg 421 atgccaccta cggcaagctg accctgaagc tcatctgcac caccggcaag ctgcccgtgc 481 cctggcccac cctcgtgacc accctgggct acggcctgca gtgcttcgcc cgctaccccg 541 accacatgaa gcagcacgac ttcttcaagt ccgccatgcc cgaaggctac gtccaggagc 601 gcaccatctt cttcaaggac gacggcaact acaagacccg cgccgaggtg aagttcgagg 661 gcgacaccct ggtgaaccgc atcgagctga agggcatcga cttcaaggag gacggcaaca 721 tcctggggca caagctggag tacaactaca acagccacaa cgtctatatc accgccgaca 781 agcagaagaa cggcatcaag gccaacttca agatccgcca caacatcgag gacggcggcg 841 tgcagctcgc cgaccactac cagcagaaca cccccatcgg cgacggcccc gtgctgctgc 901 ccgacaacca ctacctgagc taccagtccg ccctgagcaa agaccccaac gagaagcgcg 961 atcacatggt cctgctggag ttcgtgaccg ccgccgggat cactctcggc atggacgagc 1021 tgtacaagta accatggcta gatcataatc agccatacca catttgtaga ggttttactt 1081 gctttaaaaa acctcccaca cctccccctg aacctgaaac ataaaatgaa tgcaattgtt 1141 gttgttaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat 1201 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat 1261 gtatcgaatt cgtacgcgcc cggggagccc aagggcacgc cctggcaccc ggcgcgccag 1321 agggcagagg cagcctgctc acctgcggcg acgtggaaga gaaccctggg cctatgaagg 1381 ctctcacggc ccgacaacag gaagtttttg atttgatacg ggatcatata tcccaaacgg 1441 gtatgcctcc gacccgcgca gagatagcac agcgactggg ctttcgatcg cctaacgccg 1501 cggaggagca cttgaaggca ctggcccgca agggtgtcat tgaaatcgtg tccggtgcga 1561 gccgcggaat ccggctgttg caggaggagg aagagggcct gccactggtg ggacgcgtgg 1621 ccgctggcga gccgctgctg gcccagcaac acatagaggg acactatcag gtggacccct 1681 ccttgtttaa gccaaatgct gatttcctgt tgcgggtgtc gggaatgtcc atgaaggaca 1741 tcggtattat ggatggtgac ctcctcgccg tccataagac acaggatgtc aggaacggcc 1801 aggtagtcgt tgccaggata gacgatgagg tcactgtgaa acgtctcaag aagcaaggca 1861 ataaggtcga gctgctgccg gagaatagcg agttcaagcc gatcgtggtg gatctgcgac 1921 agcagtcctt tactatcgag ggcttggccg tgggtgtgat ccgcaacgga gattggctgg 1981 gatccacgcc gatggagttc cagtacctgc ccgatacgga tgaccgtcac cgtatcgaag 2041 aaaagcggaa gcgaacctat gaaaccttca agtccatcat gaaaaagtcc cccttctcgg 2101 gccccacgga cccgcgcccc ccgccccgtc gtattgcggt tccttcgcgc agcagtgcca 2161 gcgtccccaa acccgcaccg cagccctacc cgttcacttc ctcccttagc acgattaact 2221 atgatgagtt ccccacgatg gtgttcccca gtggacaaat ttcccaggca tcggcactgg 2281 ctccggcccc accgcaagtc ctcccccagg cgcccgctcc ggcaccggct cccgcaatgg 2341 tgagtgctct ggcccaggcc cccgctccag tccccgtgct ggcgcctgga cccccacagg 2401 cagttgcccc tcctgctccg aaaccaacgc aggcgggcga aggaaccctg agcgaggccc 2461 tcttgcagct tcagttcgat gacgaagact tgggagccct gctgggtaac agcacagacc 2521 ctgccgtatt caccgatctc gcatccgtgg acaacagcga gtttcagcag ctcttgaatc 2581 agggaatccc ggtcgcacct cataccacag agcccatgct gatggaatac ccggaggcta 2641 tcacgcgact ggtgaccggc gcacagcgac cacccgatcc agcccctgcc ccactgggtg 2701 ccccgggttt gcccaatggc ctcctcagcg gcgatgagga tttctccagc atcgctgata 2761 tggatttctc cgctttgctg agccagataa gctcctaagc ggccgcataa cttcgtatag 2821 catacattat acgaagttat accggtggcc gcgaagttcc tattcttcaa atagtatagg 2881 aacttcgagc tcccggagag atgcagaagc tttgcgtact cgcaaattat taaaaataaa 2941 actttaaaaa taatttcgtc taattaatat tatgagttaa ttcaaacccc acggacatgc 3001 taagggttaa tcaacaatca tatcgctgtc tcactcagac tcaatacgac actcagaata 3061 ctattccttt cactcgcact tattgcaagc atacgttaag tggatgtctc ttgccgacgg 3121 gaccacctta tgttatttca tcatggtctg gccattctca tcgtgagctt ccgggtgctc 3181 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 3241 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 3301 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 3361 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 3421 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 3481 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 3541 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 3601 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 3661 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 3721 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 3781 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 3841 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 3901 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 3961 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4021 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4081 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4141 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4201 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 4261 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 4321 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 4381 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 4441 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 4501 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 4561 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 4621 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 4681 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 4741 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 4801 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 4861 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 4921 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 4981 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5041 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5101 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5161 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 5221 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 5281 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 5341 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 5401 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 5461 taaagggagc ccccgattta gagcttgacg gggaaagccg gccttctccg ccgccgatct 5521 gccgctggac tacgtgggtc tggcccatga tgaaataaca taaggtggtc ccgtcgatag 5581 ccgaagctta ccgaagtata cacttaaatt cagtgcacgt ttgcttgttg agaggaaagg 5641 ttgtgtgcgg acgaattttt ttttgaaaac attaaccctt acgtggaata aaaaaaaatg 5701 aaatattgca aattttgctg caaagctgtg actggagtaa aattaattca cgtgccgaag 5761 tgtgctatta agagaaaatt gtgggagcag agccttgggt gcagccttgg tgaaaactcc 5821 caaatttgtg atacccactt taatgattcg cagtggaagg ctgcacctgc aaaaggtcag 5881 acatttaaaa ggaggcgact caacgcagat gccgtaccta gtaaagtgat agagcctgaa 5941 ccagaaaaga taaaagaagg ctataccagt gggagtacac aaacagagta agtttgaata 6001 gtaaaaaaaa tcatttatgt aaacaataac gtgactgtgc gttaggtcct gttcattgtt 6061 taatgaaaat aagagcttga gggaaaaaat tcgtactttg gagtacgaaa tgcgtcgttt 6121 agagcagcag c //