LOCUS Exported 6434 bp ds-DNA circular SYN 24-JUN-2019 DEFINITION synthetic circular DNA ACCESSION . VERSION . KEYWORDS . SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6434) AUTHORS Eric Schreiter TITLE Direct Submission JOURNAL Exported Wednesday, Jun 26, 2019 from SnapGene Viewer 4.3.10 https://www.snapgene.com FEATURES Location/Qualifiers source 1..6434 /organism="synthetic DNA construct" /mol_type="other DNA" promoter 256..703 /label=hSyn promoter /note="human synapsin I promoter; confers neuron-specific expression (Kugler et al., 2003)" protein_bind complement(733..766) /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites." protein_bind complement(829..862) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." CDS complement(873..2726) /label=Positron-ST CDS complement(876..1070) /codon_start=1 /label=Kir2.1_proximal restriction and clustering signal /translation="QSQPILNTKEMAPQSKPPEELEMSSMPSPVAPLPARTEGVIDMRS MSSIDSFISCATDFPEATRF" CDS complement(1071..1091) /codon_start=1 /label=ER export sequence from Kir2.1 /translation="FCYENEV" CDS complement(1092..1151) /codon_start=1 /label=TS from Kir2.1 /translation="KSRITSEGEYIPLDQIDINV" CDS complement(1152..2045) /codon_start=1 /label=HaloI4 /translation="IGTGFPFDPHYVEVLGERMHYVDVGPRDGTPVLFLHGNPTSSYVW RNIIPHVAPTHRCIAPDLIGMGKSDKPDLGYFFDDHVRFMDAFIEALGLEEVVLVIHDW GSALGFHWAKRNPERVKGIAFMEFIHPIPTWDEWPEFARETFQAFRTTDVGRKLIIDQN VFIEGTLPMGVVRPLTEVEMDHYREPFLNPVDREPLWRFPNELPIAGEPANIVALVEEY MDWLHQSPVPKLLFWGTPGVLIPPAEAARLAKSLPNCKAVDIGPGLNLLQEDNPDLIGS EIARWLSTLEISGEPTT" CDS complement(2046..2726) /codon_start=1 /label=Ace2_D92N_E199V /translation="MADVETETGMIAQWIVFAIMAAAAIAFGVAVHFRPSELKSAYYIN IAICTIAATAYYAMAVNYQDLTMNGERQVVYARYIDWVLTTPLLLLNLIVMTKMGGVMI SWVIGADIFMIVFGILGAFEDEHKFKWVYFIAGCVMQAVLTYGMYNATWKDDLKKSPEY HSSYVSLLVFLSILWVFYPVVWAFGSGSGVLSVDNVAILMGILDVLAKPLFGMGCLIAH ETIFK" regulatory 2723..2732 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" protein_bind 2743..2776 /label=lox2272 /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GGATACTT). lox2272 sites are compatible with each other, but incompatible with loxP or loxN sites." protein_bind 2847..2880 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." misc_feature 2922..3510 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" CDS complement(3393..3404) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" polyA_signal 3553..3674 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" rep_origin complement(4202..4790) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" CDS complement(4961..5821) /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" promoter complement(5822..5926) /gene="bla" /label=AmpR promoter rep_origin complement(5952..6407) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" ORIGIN 1 gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctgcgc gctcgctcgc 61 tcactgaggc cgcccgggca aagcccgggc gtcgggcgac ctttggtcgc ccggcctcag 121 tgagcgagcg agcgcgcaga gagggagtgg ccaactccat cactaggggt tccttgtagt 181 taatgattaa cccgccatgc tacttatcta cgtagccatg ctctaggaag atctctgcag 241 agggccctgc gtatgagtgc aagtgggttt taggaccagg atgaggcggg gtgggggtgc 301 ctacctgacg accgaccccg acccactgga caagcaccca acccccattc cccaaattgc 361 gcatccccta tcagagaggg ggaggggaaa caggatgcgg cgaggcgcgt gcgcactgcc 421 agcttcagca ccgcggacag tgccttcgcc cccgcctggc ggcgcgcgcc accgccgcct 481 cagcactgaa ggcgcgctga cgtcactcgc cggtcccccg caaactcccc ttcccggcca 541 ccttggtcgc gtccgcgccg ccgccggccc agccggaccg caccacgcga ggcgcgagat 601 aggggggcac gggcgcgacc atctgcgctg cggcgccggc gactcagcgc tgcctcagtc 661 tgcggtgggc agcggaggag tcgtgtcgtg cctgagagcg cagtcgaatt caagctgcta 721 gcaaggatcc ccataacttc gtataaagta tcctatacga agttatatca aaataggaag 781 accaatgctt caccatcgac ccgaattgcc aagcatcacc atcgacccat aacttcgtat 841 aatgtatgct atacgaagtt atactagagc tttcagaatc gagtagcctc ggggaaatct 901 gttgcgcaag agatgaagct atcaatagat gacatactcc tcatgtcaat cactccttca 961 gtcctggcag ggaggggagc cacgggactt ggcatactgg acatctcaag ctcttcggga 1021 ggcttagact gtggagccat ttcctttgtg ttcaggatag gctgactttg cacctcgttc 1081 tcgtagcaga acacgttgat gtcgatctgg tccaggggga tgtactcgcc ctcgctggtg 1141 atcctgctct tagtggttgg ctcgccggaa atctcgagcg tcgacagcca gcgcgcgatc 1201 tcgctgccga tcaggtccgg gttgtcttct tgcagcagat tcagacccgg gccgatgtcc 1261 acagccttgc agttaggcag gcttttggcc aggcgagcgg cttcggccgg tgggatcaga 1321 acgcctgggg tgccccagaa cagcagcttc gggacagggg actggtgcag ccagtccatg 1381 tattcttcga ccagcgcgac gatgttcgct ggctcaccgg cgattggcag ctcgtttggg 1441 aagcgccaca gtggctcgcg gtcaacagga ttcaggaacg gctcgcggta atggtccatc 1501 tcgacttcag tcagcgggcg gacgacaccc atcggcagcg taccctcgat aaaaacgttc 1561 tgatcgatga tcagcttgcg gccgacgtcg gtggtgcgga aggcctggaa ggtctcgcgg 1621 gcaaattctg gccattcgtc ccaggtcggg atagggtgga tgaactccat aaatgcaata 1681 cctttgacgc gctctggatt gcgcttggcc cagtggaaac ccagagcgga gccccagtcg 1741 tgaatgacca ggacgacctc ttccagaccc agggcttcga tgaaggcatc catgaagcgg 1801 acgtggtcgt cgaagaaata acccaggtct ggtttgtcgg atttgcccat accgatcagg 1861 tctggagcaa tgcagcgatg ggtcggtgca acatgcggga tgatgttgcg ccacacgtag 1921 gaggaggtcg ggttaccgtg caggaacagc acaggggtgc catcgcgcgg accaacatcg 1981 acgtagtgca tgcgctcgcc caggacttcc acataatggg ggtcgaatgg aaagccggta 2041 ccgatcttga agatagtctc atgggcaatg aggcacccca ttccaaacag tggcttagcg 2101 agcacatcca ggattcccat gagaatggcc acattgtcga cggacagcac gccactacca 2161 gacccgaaag cccacacgac aggataaaac acccagagga ttgacaggaa gacgagcaga 2221 ctgacatagg agctatggta ctcggggctt ttcttcagat cgtctttcca agtggcgtta 2281 tacatcccgt atgtcaggac tgcctgcatc acacatccag cgataaagta cacccatttg 2341 aacttgtgtt catcctcgaa ggcgcccaga ataccaaaca cgatcatgaa aatgtctgcg 2401 ccgatgaccc aagaaatcat cactccgccc atcttggtca tgacgatgag gttgagcagg 2461 agcagtggtg tggtcagcac ccagtcaata tatcttgcgt agaccacctg cctttcacca 2521 ttcattgtca ggtcctggta gttcacggcc attgcatagt aagcggtagc ggcgatagtg 2581 cagatggcaa tgttgatata gtatgcgctc ttcagctctg aaggccgaaa gtgcacagcc 2641 actccaaaag caatagcagc agcagccata atagcaaaga caatccactg tgcaatcata 2701 ccggtctcgg tttccacgtc agccatggtg gcggatccta gtataacttc gtataggata 2761 ctttatacga agttatcatt gggattcttc ctattttgat ccaagcatca ccatcgaccc 2821 tctagtccag atctcaccat cgacccataa cttcgtatag catacattat acgaagttat 2881 gtccctcgaa gaggttcgaa ttcgatatca agcttatcga taatcaacct ctggattaca 2941 aaatttgtga aagattgact ggtattctta actatgttgc tccttttacg ctatgtggat 3001 acgctgcttt aatgcctttg tatcatgcta ttgcttcccg tatggctttc attttctcct 3061 ccttgtataa atcctggttg ctgtctcttt atgaggagtt gtggcccgtt gtcaggcaac 3121 gtggcgtggt gtgcactgtg tttgctgacg caacccccac tggttggggc attgccacca 3181 cctgtcagct cctttccggg actttcgctt tccccctccc tattgccacg gcggaactca 3241 tcgccgcctg ccttgcccgc tgctggacag gggctcggct gttgggcact gacaattccg 3301 tggtgttgtc ggggaaatca tcgtcctttc cttggctgct cgcctgtgtt gccacctgga 3361 ttctgcgcgg gacgtccttc tgctacgtcc cttcggccct caatccagcg gaccttcctt 3421 cccgcggcct gctgccggct ctgcggcctc ttccgcgtct tcgccttcgc cctcagacga 3481 gtcggatctc cctttgggcc gcctccccgc atcgataccg tcgacccggg cggccgcttc 3541 gagcagacat gataagatac attgatgagt ttggacaaac cacaactaga atgcagtgaa 3601 aaaaatgctt tatttgtgaa atttgtgatg ctattgcttt atttgtaacc attataagct 3661 gcaataaaca agttaacaac aacaattgca ttcattttat gtttcaggtt cagggggaga 3721 tgtgggaggt tttttaaagc aagtaaaacc tctacaaatg tggtaaaatc gataaggatc 3781 ttcctagagc atggctacgt agataagtag catggcgggt taatcattaa ctacaaggaa 3841 cccctagtga tggagttggc cactccctct ctgcgcgctc gctcgctcac tgaggccggg 3901 cgaccaaagg tcgcccgacg cccgggcttt gcccgggcgg cctcagtgag cgagcgagcg 3961 cgcagctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct 4021 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat 4081 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga 4141 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt 4201 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt 4261 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc 4321 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa 4381 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct 4441 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta 4501 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg 4561 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc 4621 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta 4681 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg 4741 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt 4801 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg 4861 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta 4921 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg 4981 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg 5041 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc 5101 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg 5161 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg 5221 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag 5281 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat 5341 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc 5401 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc 5461 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa 5521 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac 5581 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt 5641 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc 5701 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa 5761 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca 5821 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat 5881 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa 5941 aagtgccacc taaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta 6001 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga 6061 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa 6121 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga 6181 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc 6241 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga 6301 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg 6361 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtccc attcgccatt 6421 caggctgcgc aact //