LOCUS Exported 7660 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Express NMNAT2. ACCESSION . VERSION . KEYWORDS AV4-EF1-NMNAT2 SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7660) TITLE 22027994 REFERENCE 2 (bases 1 to 7660) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7660 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 187..1365 /label=EF-1-alpha promoter /note="strong constitutive promoter for human elongation factor EF-1-alpha" intron 418..1356 /label=EF-1-alpha intron A /note="intron upstream of the start codon of human EF-1-alpha" primer_bind 1313..1333 /label=EF1a-F /note="Human elongation factor-1a promoter, forward primer" CDS 1387..1413 /codon_start=1 /product="HA (human influenza hemagglutinin) epitope tag" /label=HA /translation="YPYDVPDYA" misc_feature 2353..2926 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" misc_feature 2376..2928 /label=IRES /note="internal ribosome entry site (IRES) of the encephalomyocarditis virus (EMCV)" primer_bind complement(2520..2537) /label=IRES reverse /note="IRES internal ribosome entry site, reverse primer. Also called pCDH-rev" primer_bind 2747..2766 /label=IRES-F /note="IRES internal ribosome entry site, forward primer" CDS 2946..3665 /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 2946..3662 /codon_start=1 /product="enhanced GFP" /label=EGFP /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL EFVTAAGITLGMDELYK" primer_bind complement(2991..3012) /label=EGFP-N /note="EGFP, reverse primer" primer_bind complement(3252..3271) /label=EXFP-R /note="For distinguishing EGFP variants, reverse primer" primer_bind 3599..3620 /label=EGFP-C /note="EGFP, forward primer" primer_bind complement(3771..3790) /label=Bglob-pA-R /note="Rabbit beta-globin polyA region, reverse primer" polyA_signal 3836..3891 /label=-beta-globin poly(A) signal /note="rabbit beta-globin polyadenylation signal (Gil and Proudfoot, 1987)" primer_bind complement(3890..3909) /label=rbglobpA-R /note="Rabbit beta-globin polyA, reverse primer. Also called rb-glob-pA-term-R" repeat_region 4301..4430 /label=AAV2 ITR promoter complement(4501..4519) /label=SP6 promoter /note="promoter for bacteriophage SP6 RNA polymerase" primer_bind complement(4502..4519) /label=SP6 /note="SP6 promoter, forward primer" primer_bind complement(4537..4553) /label=M13 rev /note="common sequencing primer, one of multiple similar variants" primer_bind complement(4537..4553) /label=M13 Reverse /note="In lacZ gene. Also called M13-rev" primer_bind complement(4550..4572) /label=M13/pUC Reverse /note="In lacZ gene" protein_bind 4561..4577 /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(4585..4615) /label=lac promoter /note="promoter for the E. coli lac operon" protein_bind 4630..4651 /label=CAP binding site /bound_moiety="E. coli catabolite activator protein" /note="CAP binding activates transcription in the presence of cAMP." primer_bind complement(4768..4785) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4939..5527) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5019..5038) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5698..6558) /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 6321..6340 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6559..6663) /gene="bla" /label=AmpR promoter rep_origin complement(6741..7196) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(6878..6899) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7090..7109 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7322..7344 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7336..7353 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7337..7353 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" primer_bind 7360..7379 /label=T7 /note="T7 promoter, forward primer" promoter 7360..7378 /label=T7 promoter /note="promoter for bacteriophage T7 RNA polymerase" ORIGIN 1 acgtgagtgt tttgcgacat tttgcgacac catgtggtca cgctgggtat ttaagcccga 61 gtgagcacgc agggtctcca tacgcgccct gcagtttccc aagctttgca aagatggata 121 aagttttaaa cagagaggaa tctttgcagc taatggacct tctaggtctt gaaaggagtg 181 ggaattggct ccggtgcccg tcagtgggca gagcgcacat cgcccacagt ccccgagaag 241 ttggggggag gggtcggcaa ttgaaccggt gcctagagaa ggtggcgcgg ggtaaactgg 301 gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg gtgggggaga accgtatata 361 agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt ttgccgccag aacacaggta 421 agtgccgtgt gtggttcccg cgggcctggc ctctttacgg gttatggccc ttgcgtgcct 481 tgaattactt ccacctggct gcagtacgtg attcttgatc ccgagcttcg ggttggaagt 541 gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt cgcctcgtgc ttgagttgag 601 gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt ggcaccttcg cgcctgtctc 661 gctgctttcg ataagtctct agccatttaa aatttttgat gacctgctgc gacgcttttt 721 ttctggcaag atagtcttgt aaatgcgggc caagatctgc acactggtat ttcggttttt 781 ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca catgttcggc gaggcggggc 841 ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc aagctggccg gcctgctctg 901 gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg cggcaaggct ggcccggtcg 961 gcaccagttg cgtgagcgga aagatggccg cttcccggcc ctgctgcagg gagctcaaaa 1021 tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac ccacacaaag gaaaagggcc 1081 tttccgtcct cagccgtcgc ttcatgtgac tccacggagt accgggcgcc gtccaggcac 1141 ctcgattagt tctcgagctt ttggagtacg tcgtctttag gttgggggga ggggttttat 1201 gcgatggagt ttccccacac tgagtgggtg gagactgaag ttaggccagc ttggcacttg 1261 atgtaattct ccttggaatt tgcccttttt gagtttggat cttggttcat tctcaagcct 1321 cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt cgtgaggaat tggatccacc 1381 atggcatacc catatgatgt ccccgactac gcgaccgaga ccaccaagac ccacgttatc 1441 ttgctcgcct gcggcagctt caatcccatc accaaagggc acattcagat gtttgaaaga 1501 gccagggatt atctgcacaa aactggaagg tttattgtga ttggcgggat tgtctcccct 1561 gtccacgact cctatggaaa acagggcctc gtgtcaagcc ggcaccgtct catcatgtgt 1621 cagctggccg tccagaattc tgattggatc agggtggacc cttgggagtg ctaccaggac 1681 acctggcaga cgacctgcag cgtgttggaa caccaccggg acctcatgaa gagggtgact 1741 ggctgcatcc tctccaatgt caacacacct tccatgacac ctgtgatcgg acagccacaa 1801 aacgagaccc cccagcccat ttaccagaac agcaacgtgg ccaccaagcc cactgcagcc 1861 aagatcttgg ggaaggtggg agaaagcctc agccggatct gctgtgtccg cccgccggtg 1921 gagcgtttca cctttgtaga tgagaatgcc aatctgggca cggtgatgcg gtatgaagag 1981 attgagctac ggatcctgct gctgtgtggt agtgacctgc tggagtcctt ctgcatccca 2041 gggctctgga acgaggcaga tatggaggtg attgttggtg actttgggat tgtggtggtg 2101 ccccgggatg cagccgacac agaccgaatc atgaatcact cctcaatact ccgcaaatac 2161 aaaaacaaca tcatggtggt gaaggatgac atcaaccatc ccatgtctgt tgtcagctca 2221 accaagagca ggctggccct gcagcatggg gacggccatg ttgtggatta cctgtcccag 2281 ccggtcatcg actacatcct caaaagccag ctgtacatca atgcctccgg ctaggcggcc 2341 gcaattgatc cgcccctctc cctccccccc ccctaacgtt actggccgaa gccgcttgga 2401 ataaggccgg tgtgcgtttg tctatatgtt attttccacc atattgccgt cttttggcaa 2461 tgtgagggcc cggaaacctg gccctgtctt cttgacgagc attcctaggg gtctttcccc 2521 tctcgccaaa ggaatgcaag gtctgttgaa tgtcgtgaag gaagcagttc ctctggaagc 2581 ttcttgaaga caaacaacgt ctgtagcgac cctttgcagg cagcggaacc ccccacctgg 2641 cgacaggtgc ctctgcggcc aaaagccacg tgtataagat acacctgcaa aggcggcaca 2701 accccagtgc cacgttgtga gttggatagt tgtggaaaga gtcaaatggc tctcctcaag 2761 cgtattcaac aaggggctga aggatgccca gaaggtaccc cattgtatgg gatctgatct 2821 ggggcctcgg tacacatgct ttacatgtgt ttagtcgagg ttaaaaaaac gtctaggccc 2881 cccgaaccac ggggacgtgg ttttcctttg aaaaacacga tgataatatg gccacaagat 2941 ccaccatggt gagcaagggc gaggagctgt tcaccggggt ggtgcccatc ctggtcgagc 3001 tggacggcga cgtaaacggc cacaagttca gcgtgtccgg cgagggcgag ggcgatgcca 3061 cctacggcaa gctgaccctg aagttcatct gcaccaccgg caagctgccc gtgccctggc 3121 ccaccctcgt gaccaccctg acctacggcg tgcagtgctt cagccgctac cccgaccaca 3181 tgaagcagca cgacttcttc aagtccgcca tgcccgaagg ctacgtccag gagcgcacca 3241 tcttcttcaa ggacgacggc aactacaaga cccgcgccga ggtgaagttc gagggcgaca 3301 ccctggtgaa ccgcatcgag ctgaagggca tcgacttcaa ggaggacggc aacatcctgg 3361 ggcacaagct ggagtacaac tacaacagcc acaacgtcta tatcatggcc gacaagcaga 3421 agaacggcat caaggtgaac ttcaagatcc gccacaacat cgaggacggc agcgtgcagc 3481 tcgccgacca ctaccagcag aacaccccca tcggcgacgg ccccgtgctg ctgcccgaca 3541 accactacct gagcacccag tccgccctga gcaaagaccc caacgagaag cgcgatcaca 3601 tggtcctgct ggagttcgtg accgccgccg ggatcactct cggcatggac gagctgtaca 3661 agtaagaatt catcgatcta gagcatgcaa ttcactcctc aggtgcaggc tgcctatcag 3721 aaggtggtgg ctggtgtggc caatgccctg gctcacaaat accactgaga tctttttccc 3781 tctgccaaaa attatgggga catcatgaag ccccttgagc atctgacttc tggctaataa 3841 aggaaattta ttttcattgc aatagtgtgt tggaattttt tgtgtctctc actcggaagg 3901 acatatggga gggcaaatca tttaaaacat cagaatgagt atttggttta gagtttggca 3961 acatatgccc atatgctggc tgccatgaac aaaggttggc tataaagagg tcatcagtat 4021 atgaaacagc cccctgctgt ccattcctta ttccatagaa aagccttgac ttgaggttag 4081 atttttttta tattttgttt tgtgttattt ttttctttaa catccctaaa attttcctta 4141 catgttttac tagccagatt tttcctcctc tcctgactac tcccagtcat agctgtccct 4201 cttctcttat ggagatccct cgacctgcag cccaagctta ggcgcgttct agagcatggc 4261 tacgtagata agtagcatgg cgggttaatc attaactaca aggaacccct agtgatggag 4321 ttggccactc cctctctgcg cgctcgctcg ctcactgagg ccgggcgacc aaaggtcgcc 4381 cgacgcccgg gctttgcccg ggcggcctca gtgagcgagc gagcgcgcag ggttcgaaat 4441 cgataagctt ggatcgatcc ggagagctcg tcgacggcgc gccatgcata gcttgagtat 4501 tctatagtgt cacctaaata gcttggcgta atcatggtca tagctgtttc ctgtgtgaaa 4561 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg 4621 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca 4681 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg 4741 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg 4801 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg 4861 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa 4921 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg 4981 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc 5041 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc 5101 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc 5161 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg 5221 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc 5281 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga 5341 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc 5401 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac 5461 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg 5521 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc 5581 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa 5641 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta 5701 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt 5761 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag 5821 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca 5881 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc 5941 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt 6001 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag 6061 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt 6121 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat 6181 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt 6241 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc 6301 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat 6361 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag 6421 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt 6481 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg 6541 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta 6601 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc 6661 gcgcacattt ccccgaaaag tgccacctga tgcggtgtga aataccgcac agatgcgtaa 6721 ggagaaaata ccgcatcagg aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa 6781 tttttgttaa atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa 6841 atcaaaagaa tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact 6901 attaaagaac gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc 6961 actacgtgaa ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa 7021 tcggaaccct aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc 7081 gagaaaggaa gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt 7141 cacgctgcgc gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccat 7201 tcgccattca ggctgcgcaa ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta 7261 cgccagctgg cgaaaggggg atgtgctgca aggcgattaa gttgggtaac gccagggttt 7321 tcccagtcac gacgttgtaa aacgacggcc agtgaattgt aatacgactc actatagggc 7381 gaattgggcc cggcgcgccg acgcgcatgc tcctctagct agactcgagg aattcggtac 7441 cccctgcgcg ctcgctcgct cactgaggcc gcccgggcaa agcccgggcg gcctcagtga 7501 gcgagcgagc gcgcagagag ggagtggcca actccatcac taggggttcc ttgtagttaa 7561 tgattaaccc gccatgctac ttatctacgt agccatgctc tagagtggag tcgtgacgtg 7621 aattacgtca tagggttagg gaggtcctgt attagaggtc //