LOCUS Exported 6918 bp ds-DNA circular SYN 30-JAN-2018 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS Gncp-iGluSnFR SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6918) AUTHORS . TITLE Direct Submission JOURNAL Exported Thursday, February 1, 2018 from SnapGene Viewer 3.2.1 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6918 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 466..845 /note="CMV enhancer" /note="human cytomegalovirus immediate early enhancer" promoter 846..1049 /note="CMV promoter" /note="human cytomegalovirus (CMV) immediate early promoter" promoter 1094..1112 /note="T7 promoter" /note="promoter for bacteriophage T7 RNA polymerase" CDS 1193..1255 /codon_start=1 /product="leader sequence from mouse immunoglobulin kappa light chain" /note="Ig-kappa leader" /translation="METDTLLLWVLLLWVPGSTGD" misc_feature 1262..2911 /note="Gncp-iGluSnFR" CDS 1787..1804 /codon_start=1 /product="6xHis affinity tag" /note="6xHis" /translation="HHHHHH" CDS 1808..1840 /codon_start=1 /product="leader peptide from bacteriophage T7 gene 10" /note="T7 tag (gene 10 leader)" /note="promotes efficient translation in E. coli" /translation="MASMTGGQQMG" CDS 1844..1867 /codon_start=1 /product="Xpress(TM) epitope tag, including an enterokinase recognition and cleavage site" /note="Xpress(TM) tag" /translation="DLYDDDDK" CDS 2924..2953 /codon_start=1 /product="Myc (human c-Myc oncogene) epitope tag" /note="Myc" /translation="EQKLISEEDL" polyA_signal 3131..3355 /note="bGH poly(A) signal" /note="bovine growth hormone polyadenylation signal" rep_origin complement(3487..4075) /direction=LEFT /note="ori" /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" polyA_signal 4404..4451 /note="HSV TK poly(A) signal" /note="herpes simplex virus thymidine kinase polyadenylation signal (Cole and Stacy, 1985)" CDS complement(4683..5477) /codon_start=1 /gene="aph(3')-II (or nptII)" /product="aminoglycoside phosphotransferase from Tn5" /note="NeoR/KanR" /note="confers resistance to neomycin, kanamycin, and G418 (Geneticin(R))" /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF" CDS complement(5925..6785) /codon_start=1 /gene="bla" /product="beta-lactamase" /note="AmpR" /note="confers resistance to ampicillin, carbenicillin, and related antibiotics" /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS LIKHW" promoter complement(6786..6890) /gene="bla" /note="AmpR promoter" rep_origin 6917..454 /direction=RIGHT /note="f1 ori" /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag cgtgaccgct 61 acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg 121 ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt 181 gctttacggc acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca 241 tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga 301 ctcttgttcc aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa 361 gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac 421 gcgaatttta acaaaatatt aacgcttaca atttacgcgc gcgttgacat tgattattga 481 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc 541 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat 601 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc 661 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc 721 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt 781 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta 841 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg 901 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac 961 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg 1021 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact 1081 ggcttatcga aattaatacg actcactata gggagaccca agcttggtac cgagctcgga 1141 tccactagta acggccgcca gtgtgctgga attcggcttg gggatatcca ccatggagac 1201 agacacactc ctgctatggg tactgctgct ctgggttcca ggttccactg gtgacagatc 1261 tgtgagcaag ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg 1321 cgacgtaaac ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg 1381 caagctgacc ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct 1441 cgtgaccacc ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca 1501 gcacgacttc ttcaagtccg ccatgcccga aggctacatc caggagcgca ccatcttctt 1561 caaggacgac ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt 1621 gaaccgcatc gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcataa 1681 gctggagtac aactttaaca accccctgaa catgaatttc gaactgtcag acgaaatgaa 1741 agcactgttc aaagaaccga atgacaaggc actgaaaggc ggctctcatc atcatcatca 1801 tcatggtatg gctagcatga ctggtggaca gcaaatgggt cgggatctgt acgacgatga 1861 cgataaggat cctggctcga gcggatccgc cgcaggcagc acgctggaca aaatcgccaa 1921 aaacggtgtg attgtcgtcg gtcaccgtga atcttcagtg cctttctctt attacgacaa 1981 tcagcaaaaa gtggtgggtt actcgcagga ttactccaac gccattgttg aagcagtgaa 2041 aaagaaactc aacaaaccgg acttgcaggt aaaactgatt ccgattacct cacaaaaccg 2101 tattccactg ctgcaaaacg gcactttcga ttttgaatgt ggttctacca ccaacaacgt 2161 cgaacgccaa aaacaggcgg ctttctctga cactattttc gtggtcggta cgcgcctgtt 2221 gaccaaaaag ggtggcgata tcaaagattt tgccaacctg aaagacaaag ccgtagtcgt 2281 cacttccggc actacctctg aagttttgct caacaaactg aatgaagagc aaaaaatgaa 2341 tatgcgcatc atcagcgcca aagatcacgg tgactctttc cgcaccctgg aaagcggtcg 2401 tgccgttgcc tttatgatgg atgacgtgct gctggccggt gaacgtgcga aagcgaagaa 2461 accagacaac tgggaaatcg tcggcaagcc gcagtctcag gaggcctacg gttgtatgtt 2521 gcgtaaagat gatccgcagt tcaaaaagct gatggatgac accatcgctc aggtgcagac 2581 ctccggtgaa gcggaaaaat ggtttgataa gtggttcaaa aatccaattc tggtgagcca 2641 caacgtctat atcatggccg acaagcagag gaacggcatc aaggcgaact tcaagatccg 2701 ccacaacatc gaggacggcg gcgtgcagct cgcctaccac taccagcaga acacccccat 2761 cggcgacggc cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccaaactttc 2821 gaaagacccc aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg 2881 gatcactctc ggcatggacg agctgtacaa gctgcaggtc gacgaacaaa aactcatctc 2941 agaagaggat ctgaatgctg tgggccagga cacgcaggag gtcatcgtgg tgccacactc 3001 cttgcccttt aaggtggtgg tgatctcagc catcctggcc ctggtggtgc tcaccatcat 3061 ctcccttatc atcctcatca tgctttggca gaagaagcca cgttaggcgg ccgctcgaga 3121 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct 3181 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca 3241 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag 3301 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct 3361 gaggcggaaa gaaccagtgg cggtaatacg gttatccaca gaatcagggg ataacgcagg 3421 aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct 3481 ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca 3541 gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct 3601 cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc 3661 gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt 3721 tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc 3781 cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc 3841 cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg 3901 gtggcctaac tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc 3961 agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag 4021 cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga 4081 tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat 4141 tttggtcatg agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag 4201 ttttaaatca atctaaagta tatatgagta acctgaggct atggcagggc ctgccgcccc 4261 gacgttggct gcgagccctg ggccttcacc cgaacttggg gggtggggtg gggaaaagga 4321 agaaacgcgg gcgtattggc cccaatgggg tctcggtggg gtatcgacag agtgccagcc 4381 ctgggaccga accccgcgtt tatgaacaaa cgacccaaca ccgtgcgttt tattctgtct 4441 ttttattgcc gtcatagcgc gggttccttc cggtattgtc tccttccgtg tttcagttag 4501 cctcccccta gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca 4561 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc 4621 gaaatctcgt gatggcaggt tgggcgtcgc ttggtcggtc atttcgaacc ccagagtccc 4681 gctcagaaga actcgtcaag aaggcgatag aaggcgatgc gctgcgaatc gggagcggcg 4741 ataccgtaaa gcacgaggaa gcggtcagcc cattcgccgc caagctcttc agcaatatca 4801 cgggtagcca acgctatgtc ctgatagcgg tccgccacac ccagccggcc acagtcgatg 4861 aatccagaaa agcggccatt ttccaccatg atattcggca agcaggcatc gccatgggtc 4921 acgacgagat cctcgccgtc gggcatgctc gccttgagcc tggcgaacag ttcggctggc 4981 gcgagcccct gatgctcttc gtccagatca tcctgatcga caagaccggc ttccatccga 5041 gtacgtgctc gctcgatgcg atgtttcgct tggtggtcga atgggcaggt agccggatca 5101 agcgtatgca gccgccgcat tgcatcagcc atgatggata ctttctcggc aggagcaagg 5161 tgagatgaca ggagatcctg ccccggcact tcgcccaata gcagccagtc ccttcccgct 5221 tcagtgacaa cgtcgagcac agctgcgcaa ggaacgcccg tcgtggccag ccacgatagc 5281 cgcgctgcct cgtcttgcag ttcattcagg gcaccggaca ggtcggtctt gacaaaaaga 5341 accgggcgcc cctgcgctga cagccggaac acggcggcat cagagcagcc gattgtctgt 5401 tgtgcccagt catagccgaa tagcctctcc acccaagcgg ccggagaacc tgcgtgcaat 5461 ccatcttgtt caatcatgcg aaacgatcct catcctgtct cttgatcgat ctttgcaaaa 5521 gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg ccgggcctcg 5581 gcctctgcat aaataaaaaa aattagtcag ccatggggcg gagaatgggc ggaactgggc 5641 ggagttaggg gcgggatggg cggagttagg ggcgggacta tggttgctga ctaattgaga 5701 tgcatgcttt gcatacttct gcctgctggg gagcctgggg actttccaca cctggttgct 5761 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca 5821 caccctaact gacacacatt ccacagctgg ttctttccgc ctcaggactc ttcctttttc 5881 aataaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc 5941 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc 6001 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata 6061 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg 6121 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc 6181 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct 6241 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa 6301 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt 6361 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca 6421 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac 6481 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca 6541 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt 6601 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc 6661 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca 6721 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata 6781 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc 6841 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc 6901 cgaaaagtgc cacctgac //