LOCUS Exported 6537 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION transient/stable overexpression. ACCESSION . VERSION . KEYWORDS pcDNA3.1(+)/Puro/FA/GFP-N--Grb2-5 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6537) AUTHORS Gingras J, Gawor M, Bernadzki KM, Grady RM, Hallock P, Glass DJ, Sanes JR, Proszynski TJ TITLE Alpha-Dystrobrevin-1 recruits Grb2 and alpha-catulin to organize neurotransmitter receptors at the neuromuscular junction. JOURNAL J Cell Sci. 2016 Mar 1;129(5):898-911. doi: 10.1242/jcs.181180. Epub 2016 Jan 14. PUBMED 26769899 REFERENCE 2 (bases 1 to 6537) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6537 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 15..394 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 395..598 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 549..569 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 643..662 /label=T7 /note="T7 promoter, forward primer" promoter 643..661 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" CDS 715..1431 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" CDS 715..1431 /codon_start=1 /product="the original enhanced GFP (Yang et al., 1996)" /label=EGFP /note="mammalian codon-optimized" /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(760..781) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(1021..1040) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 1368..1389 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(2163..2180) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 2169..2393 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" rep_origin 2439..2867 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(2526..2545) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 2736..2757 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(2876..2896) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 2881..3210 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 3061..3196 /label=SV40 ori /note="SV40 origin of replication" primer_bind 3123..3142 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" CDS 3231..3830 /codon_start=1 /gene="pac from Streptomyces alboniger" /product="puromycin N-acetyltransferase" /label=PuroR /note="confers resistance to puromycin" /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA" primer_bind complement(3231..3250) /label=Puro-R /note="Puromycin resistance gene, reverse primer. Also called puro-variant-R" primer_bind 3727..3747 /label=Puro-F /note="Puromycin resistance gene, forward primer" polyA_signal 3990..4111 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4027..4046) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4081..4100 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4160..4176) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4160..4176) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4173..4195) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4184..4200 /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)." promoter complement(4208..4238) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4253..4274 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4391..4408) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4562..5150) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4642..4661) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5321..6181) /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 5944..5963 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6182..6286) /gene="bla" /label=AmpR promoter primer_bind complement(6361..6380) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" ORIGIN 1 cagatatacg cgttgacatt gattattgac tagttattaa tagtaatcaa ttacggggtc 61 attagttcat agcccatata tggagttccg cgttacataa cttacggtaa atggcccgcc 121 tggctgaccg cccaacgacc cccgcccatt gacgtcaata atgacgtatg ttcccatagt 181 aacgccaata gggactttcc attgacgtca atgggtggag tatttacggt aaactgccca 241 cttggcagta catcaagtgt atcatatgcc aagtacgccc cctattgacg tcaatgacgg 301 taaatggccc gcctggcatt atgcccagta catgacctta tgggactttc ctacttggca 361 gtacatctac gtattagtca tcgctattac catggtgatg cggttttggc agtacatcaa 421 tgggcgtgga tagcggtttg actcacgggg atttccaagt ctccacccca ttgacgtcaa 481 tgggagtttg ttttggcacc aaaatcaacg ggactttcca aaatgtcgta acaactccgc 541 cccattgacg caaatgggcg gtaggcgtgt acggtgggag gtctatataa gcagagctct 601 ctggctaact agagaaccca ctgcttactg gcttatcgaa attaatacga ctcactatag 661 ggagacccaa gctggctagc gtttaaactt aagcttggta ccgagctcgg atccatggtg 721 agcaagggcg aggagctgtt caccggggtg gtgcccatcc tggtcgagct ggacggcgac 781 gtaaacggcc acaagttcag cgtgtccggc gagggcgagg gcgatgccac ctacggcaag 841 ctgaccctga agttcatctg caccaccggc aagctgcccg tgccctggcc caccctcgtg 901 accaccctga cctacggcgt gcagtgcttc agccgctacc ccgaccacat gaagcagcac 961 gacttcttca agtccgccat gcccgaaggc tacgtccagg agcgcaccat cttcttcaag 1021 gacgacggca actacaagac ccgcgccgag gtgaagttcg agggcgacac cctggtgaac 1081 cgcatcgagc tgaagggcat cgacttcaag gaggacggca acatcctggg gcacaagctg 1141 gagtacaact acaacagcca caacgtctat atcatggccg acaagcagaa gaacggcatc 1201 aaggtgaact tcaagatccg ccacaacatc gaggacggca gcgtgcagct cgccgaccac 1261 taccagcaga acacccccat cggcgacggc cccgtgctgc tgcccgacaa ccactacctg 1321 agcacccagt ccgccctgag caaagacccc aacgagaagc gcgatcacat ggtcctgctg 1381 gagttcgtga ccgccgccgg gatcactctc ggcatggacg agctgtacaa gggccggcct 1441 gtggtggaat tcatggaagc catcgccaaa tatgacttca aagctactgc tgacgatgag 1501 ctgagcttca aaagggggga catccttaag gttttgaatg aagaatgtga ccagaactgg 1561 tataaggcag aactcaatgg gaaagatggc ttcatcccca agaattacat agaaatgaaa 1621 ccacatccgt ggttttttgg caaaatcccc agagccaagg cagaagaaat gctcagcaaa 1681 cagcggcatg acggggcctt cctgatccga gagagcgaga gcgctcctgg ggacttctcc 1741 ctgtccgtca agtttggaaa tgatgtgcag cacttcaagg tgctccgcga cggagccggg 1801 aagtatttcc tgtgggtggt gaagtttaat tctttgaatg agctggtaga ttaccacaga 1861 tcaacatccg tgtccaggaa ccagcagata ttcttacggg acatagaaca gatgccacag 1921 cagccaacct acgtccaggc gctctttgac tttgaccccc aggaggatgg cgagctgggc 1981 tttcgcagag gagacttcat tcatgtcatg gataactcag atcccaattg gtggaaaggg 2041 gcctgccacg ggcagaccgg catgtttccc cgcaattatg tcacccccgt gaaccggaac 2101 gtcgatatcc agcacagtgg cgcgcctcga gtctagaggg cccgtttaaa cccgctgatc 2161 agcctcgact gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc 2221 cttgaccctg gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc 2281 gcattgtctg agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg 2341 ggaggattgg gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga 2401 ggcggaaaga accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt 2461 aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc 2521 gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca 2581 agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc 2641 caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt 2701 tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac 2761 aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc 2821 ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat 2881 gtgtgtcagt tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc 2941 atgcatctca attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga 3001 agtatgcaaa gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc 3061 atcccgcccc taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt 3121 tttatttatg cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga 3181 ggcttttttg gaggcctagg cttttgcaaa aagctcccgg ggccgccacc atgaccgagt 3241 acaagcccac ggtgcgcctc gccacccgcg acgacgtccc ccgggccgta cgcaccctcg 3301 ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgacccggac cgccacatcg 3361 agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac atcggcaagg 3421 tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag agcgtcgaag 3481 cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt tcccggctgg 3541 ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag cccgcgtggt 3601 tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc agcgccgtcg 3661 tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg gagacctccg 3721 cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc gacgtcgagg 3781 tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga ttcgaaatga 3841 ccgaccaagc gacgcccaac ctgccatcac gagatttcga ttccaccgcc gccttctatg 3901 aaaggttggg cttcggaatc gttttccggg acgccggctg gatgatcctc cagcgcgggg 3961 atctcatgct ggagttcttc gcccacccca acttgtttat tgcagcttat aatggttaca 4021 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 4081 gtggtttgtc caaactcatc aatgtatctt atcatgtctg tataccgtcg acctctagct 4141 agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg aaattgttat ccgctcacaa 4201 ttccacacaa catacgagcc ggaagcataa agtgtaaagc ctggggtgcc taatgagtga 4261 gctaactcac attaattgcg ttgcgctcac tgcccgcttt ccagtcggga aacctgtcgt 4321 gccagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4381 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4441 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4501 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4561 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4621 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4681 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4741 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4801 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4861 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4921 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4981 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 5041 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 5101 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 5161 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 5221 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 5281 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 5341 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5401 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5461 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5521 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5581 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5641 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5701 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5761 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5821 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5881 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5941 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 6001 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 6061 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 6121 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 6181 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 6241 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 6301 aagtgccacc tgacgtcgac ggatcgggag atctcccgat cccctatggt gcactctcag 6361 tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt gtgtgttgga 6421 ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct tgaccgacaa 6481 ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg tacgggc //