LOCUS Exported 6770 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION tracing neuronal projection. ACCESSION . VERSION . KEYWORDS AAV-flex-plap SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 6770) TITLE Shah lab AAV-flex-plap JOURNAL Unpublished REFERENCE 2 (bases 1 to 6770) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..6770 /organism="synthetic DNA construct" /mol_type="other DNA" repeat_region 1..141 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 12..141 /label=AAV2 ITR enhancer 217..520 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 521..724 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 675..695 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 721..745 /label=LNCX /note="Human CMV promoter, forward primer" protein_bind 1358..1391 /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind 1402..1435 /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 (Lee and Saito, 1988)." CDS complement(1581..3086) /codon_start=1 /product="secreted alkaline phosphatase from human placenta" /label=SEAP /note="The natural enzyme has a C-terminal transmembrane sequence that is replaced by a GPI anchor." /translation="MLLLLLLLLGLRLQLSLGIIPVEEENPDFWNREAAEALGAAKKLQ PAQTAAKNLIIFLGDGMGVSTVTAARILKGQKKDKLGPEIPLAMDRFPYVALSKTYNVD KHVPDSGATATAYLCGVKGNFQTIGLSAAARFNQCNTTRGNEVISVMNRAKKAGKSVGV VTTTRVQHASPAGTYAHTVNRNWYSDADVPASARQEGCQDIATQLISNMDIDVILGGGR KYMFRMGTPDPEYPDDYSQGGTRLDGKNLVQEWLAKRQGARYVWNRTELMQASLDPSVT HLMGLFEPGDMKYEIHRDSTLDPSLMEMTEAALRLLSRNPRGFFLFVEGGRIDHGHHES RAYRALTETIMFDDAIERAGQLTSEEDTLSLVTADHSHVFSFGGYPLRGSSIFGLAPGK ARDRKAYTVLLYGNGPGYVLKDGARPDVTESESGSPEYRQQSAVPLDEETHAGEDVAVF ARGPQAHLVHGVQEQTFIAHVMAFAACLEPYTACDLAPPAGTTD" protein_bind complement(3111..3144) /label=loxP /bound_moiety="Cre recombinase" /note="Cre-mediated recombination occurs in the 8-bp core sequence (GCATACAT)." protein_bind complement(3155..3188) /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 (Lee and Saito, 1988)." misc_feature 3198..3786 /label=WPRE /note="woodchuck hepatitis virus posttranscriptional regulatory element" primer_bind complement(3251..3271) /label=WPRE-R /note="WPRE, reverse primer" CDS complement(3669..3680) /codon_start=1 /product="Factor Xa recognition and cleavage site" /label=Factor Xa site /translation="IEGR" primer_bind complement(3797..3814) /label=BGH-rev /note="Bovine growth hormone terminator, reverse primer. Also called BGH reverse" polyA_signal 3803..4010 /label=bGH poly(A) signal /note="bovine growth hormone polyadenylation signal" repeat_region 4033..4173 /label=AAV2 ITR /note="inverted terminal repeat of adeno-associated virus serotype 2" repeat_region 4033..4162 /label=AAV2 ITR rep_origin 4248..4703 /direction=RIGHT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(4335..4354) /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 4545..4566 /label=F1ori-F /note="F1 origin, forward primer" primer_bind complement(4720..4739) /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" primer_bind 4839..4861 /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(4899..4917) /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" promoter 4985..5089 /gene="bla" /label=AmpR promoter CDS 5090..5950 /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 complement(5308..5327) /label=Amp-R /note="Ampicillin resistance gene, reverse primer" rep_origin 6121..6709 /direction=RIGHT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind 6610..6629 /label=pBR322ori-F /note="pBR322 origin, forward primer" ORIGIN 1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc 61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca 121 actccatcac taggggttcc tgcggccgca cgcgtctagt tattaatagt aatcaattac 181 ggggtcatta gttcatagcc catatatgga gttccgcgtt acataactta cggtaaatgg 241 cccgcctggc tgaccgccca acgacccccg cccattgacg tcaataatga cgtatgttcc 301 catagtaacg ccaataggga ctttccattg acgtcaatgg gtggagtatt tacggtaaac 361 tgcccacttg gcagtacatc aagtgtatca tatgccaagt acgcccccta ttgacgtcaa 421 tgacggtaaa tggcccgcct ggcattatgc ccagtacatg accttatggg actttcctac 481 ttggcagtac atctacgtat tagtcatcgc tattaccatg gtgatgcggt tttggcagta 541 catcaatggg cgtggatagc ggtttgactc acggggattt ccaagtctcc accccattga 601 cgtcaatggg agtttgtttt ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa 661 ctccgcccca ttgacgcaaa tgggcggtag gcgtgtacgg tgggaggtct atataagcag 721 agctcgttta gtgaaccgtc agatcgcctg gagacgccat ccacgctgtt ttgacctcca 781 tagaagacac cgggaccgat ccagcctccg cggattcgaa tcccggccgg gaacggtgca 841 ttggaacgcg gattccccgt gccaagagtg acgtaagtac cgcctataga gtctataggc 901 ccacaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt 961 ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt 1021 ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat 1081 atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa 1141 tccagctacc attctgcttt tattttatgg ttgggataag gctggattat tctgagtcca 1201 agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca gctcctgggc 1261 aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattgggatt cgaacatcga 1321 tagcgacgcg tggacggtac ccttcaggtc gacgtcgata acttcgtata gcatacatta 1381 tacgaagtta tccgaatcgc aataacttcg tataggatac tttatacgaa gttatttggc 1441 gcgccaaatt taaataacgc aacgtacctt cgagggggcc caaggaattc tcagggagca 1501 gtggccgtct ccagcagcag cagggtcccg gccagcagag gaagcaacgc ggggaccacg 1561 gaccgccccg ggtgcgcggc gtcggtggtg ccggcggggg gcgccaggtc gcaggcggtg 1621 tagggctcca ggcaggcggc gaaggccatg acgtgcgcta tgaaggtctg ctcctgcacg 1681 ccgtgaacca ggtgcgcctg cgggccgcgc gcgaacaccg ccacgtcctc gcctgcgtgg 1741 gtctcttcgt ccaggggcac tgctgactgc tgccgatact cggggctccc gctctcgctc 1801 tcggtaacat ccggccgggc gccgtccttg agcacatagc ctggaccgtt tccgtatagg 1861 aggaccgtgt aggccttcct gtcccgggcc ttgccagggg ccagcccgaa gatggagctc 1921 cctcgcaggg ggtagcctcc gaaggagaag acgtgggagt ggtcggcagt gacgaggctc 1981 agcgtgtcct cctcgctggt gagctggccc gccctctcaa tggcgtcgtc gaacatgatc 2041 gtctcagtca gtgcccggta agccctgctt tcatgatgac catggtcgat gcgaccaccc 2101 tccacgaaga ggaagaagcc gcgggggttc ctgctcagca ggcgcagggc agcctctgtc 2161 atctccatca gggaggggtc cagtgtggag tctcggtgga tctcgtattt catgtctcca 2221 ggctcaaaga gacccatgag atgggtcaca gacgggtcca gggaagcctg catgagctca 2281 gtgcggttcc acacataccg ggcaccctgg cgcttcgcca gccattcctg caccagattc 2341 ttcccgtcca gcctggtccc accttggctg tagtcatctg ggtactcagg gtctggggtt 2401 cccatgcgaa acatgtactt tcggcctcca cctaggatca cgtcaatgtc catgttggag 2461 atgagctgcg tagcgatgtc ctggcacccc tcctggcggg ccgaggcagg cacgtcggcg 2521 tccgagtacc agttgcggtt caccgtgtgg gcgtaggtgc cggctggcga ggcgtgctgc 2581 actcgtgtgg tggttaccac tcccactgac ttccctgctt tcttggcccg attcatcacg 2641 gagatgacct cgttgccgcg tgtcgtgttg cactggttaa agcgggcggc tgcactcaag 2701 ccaatggtct ggaagttgcc cttgaccccg cacaggtagg ccgtggctgt ggctccactg 2761 tctggcacat gtttgtctac attgtatgtc ttggacagag ccacatatgg gaagcggtcc 2821 atggccaggg gtatctcagg ccccagtttg tccttcttct gcccttttag gatcctggca 2881 gctgtcaccg tagacacccc catcccatcg cccaggaaga tgatgaggtt cttggcggct 2941 gtctgtgcag gctgcagctt cttggcggca cccagggcct cggctgcctc gcggttccag 3001 aagtccgggt tctcctcctc aactgggatg atgcccaggg agagctgtag cctcaggccc 3061 agcagcagca gcagcagcag cagcatggtg gcaaggccgg cctcgagttg ataacttcgt 3121 ataatgtatg ctatacgaag ttatccgaat cgcaataact tcgtataaag tatcctatac 3181 gaagttatcg aaagcttaat caacctctgg attacaaaat ttgtgaaaga ttgactggta 3241 ttcttaacta tgttgctcct tttacgctat gtggatacgc tgctttaatg cctttgtatc 3301 atgctattgc ttcccgtatg gctttcattt tctcctcctt gtataaatcc tggttgctgt 3361 ctctttatga ggagttgtgg cccgttgtca ggcaacgtgg cgtggtgtgc actgtgtttg 3421 ctgacgcaac ccccactggt tggggcattg ccaccacctg tcagctcctt tccgggactt 3481 tcgctttccc cctccctatt gccacggcgg aactcatcgc cgcctgcctt gcccgctgct 3541 ggacaggggc tcggctgttg ggcactgaca attccgtggt gttgtcgggg aaatcatcgt 3601 cctttccttg gctgctcgcc tgtgttgcca cctggattct gcgcgggacg tccttctgct 3661 acgtcccttc ggccctcaat ccagcggacc ttccttcccg cggcctgctg ccggctctgc 3721 ggcctcttcc gcgtcttcgc cttcgccctc agacgagtcg gatctccctt tgggccgcct 3781 ccccgcctga ccggtgcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc 3841 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat 3901 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg 3961 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga cacgtgcgga 4021 ccgagcggcc gcaggaaccc ctagtgatgg agttggccac tccctctctg cgcgctcgct 4081 cgctcactga ggccgggcga ccaaaggtcg cccgacgccc gggctttgcc cgggcggcct 4141 cagtgagcga gcgagcgcgc agctgcctgc aggggcgcct gatgcggtat tttctcctta 4201 cgcatctgtg cggtatttca caccgcatac gtcaaagcaa ccatagtacg cgccctgtag 4261 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag 4321 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt 4381 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca 4441 cctcgacccc aaaaaacttg atttgggtga tggttcacgt agtgggccat cgccctgata 4501 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca 4561 aactggaaca acactcaacc ctatctcggg ctattctttt gatttataag ggattttgcc 4621 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattttaa 4681 caaaatatta acgtttacaa ttttatggtg cactctcagt acaatctgct ctgatgccgc 4741 atagttaagc cagccccgac acccgccaac acccgctgac gcgccctgac gggcttgtct 4801 gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca tgtgtcagag 4861 gttttcaccg tcatcaccga aacgcgcgag acgaaagggc ctcgtgatac gcctattttt 4921 ataggttaat gtcatgataa taatggtttc ttagacgtca ggtggcactt ttcggggaaa 4981 tgtgcgcgga acccctattt gtttattttt ctaaatacat tcaaatatgt atccgctcat 5041 gagacaataa ccctgataaa tgcttcaata atattgaaaa aggaagagta tgagtattca 5101 acatttccgt gtcgccctta ttcccttttt tgcggcattt tgccttcctg tttttgctca 5161 cccagaaacg ctggtgaaag taaaagatgc tgaagatcag ttgggtgcac gagtgggtta 5221 catcgaactg gatctcaaca gcggtaagat ccttgagagt tttcgccccg aagaacgttt 5281 tccaatgatg agcactttta aagttctgct atgtggcgcg gtattatccc gtattgacgc 5341 cgggcaagag caactcggtc gccgcataca ctattctcag aatgacttgg ttgagtactc 5401 accagtcaca gaaaagcatc ttacggatgg catgacagta agagaattat gcagtgctgc 5461 cataaccatg agtgataaca ctgcggccaa cttacttctg acaacgatcg gaggaccgaa 5521 ggagctaacc gcttttttgc acaacatggg ggatcatgta actcgccttg atcgttggga 5581 accggagctg aatgaagcca taccaaacga cgagcgtgac accacgatgc ctgtagcaat 5641 ggcaacaacg ttgcgcaaac tattaactgg cgaactactt actctagctt cccggcaaca 5701 attaatagac tggatggagg cggataaagt tgcaggacca cttctgcgct cggcccttcc 5761 ggctggctgg tttattgctg ataaatctgg agccggtgag cgtgggtctc gcggtatcat 5821 tgcagcactg gggccagatg gtaagccctc ccgtatcgta gttatctaca cgacggggag 5881 tcaggcaact atggatgaac gaaatagaca gatcgctgag ataggtgcct cactgattaa 5941 gcattggtaa ctgtcagacc aagtttactc atatatactt tagattgatt taaaacttca 6001 tttttaattt aaaaggatct aggtgaagat cctttttgat aatctcatga ccaaaatccc 6061 ttaacgtgag ttttcgttcc actgagcgtc agaccccgta gaaaagatca aaggatcttc 6121 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc 6181 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt 6241 cagcagagcg cagataccaa atactgtcct tctagtgtag ccgtagttag gccaccactt 6301 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc 6361 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa 6421 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac 6481 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg 6541 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga 6601 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact 6661 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa 6721 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt //