LOCUS Exported 8786 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses the Sema3E protein (truncated at cleavage site P3; ie, long), C-terminally fused to alkaline phosphatase + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Sema3e(L)-AP-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8786) AUTHORS Visser JJ, Cheng Y, Perry SC, Chastain AB, Parsa B, Masri SS, Ray TA, Kay JN, Wojtowicz WM TITLE An extracellular biochemical screen reveals that FLRTs and Unc5s mediate neuronal subtype recognition in the retina. JOURNAL Elife. 2015 Dec 3;4. pii: e08149. doi: 10.7554/eLife.08149. PUBMED 26633812 REFERENCE 2 (bases 1 to 8786) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8786 /organism="synthetic DNA construct" /mol_type="other DNA" enhancer 105..408 /label=CMV enhancer /note="human cytomegalovirus immediate early enhancer" promoter 409..612 /label=CMV promoter /note="human cytomegalovirus (CMV) immediate early promoter" primer_bind 563..583 /label=CMV-F /note="Human CMV immediate early promoter, forward primer" primer_bind 609..633 /label=LNCX /note="Human CMV promoter, forward primer" intron 748..1573 /label=CMV intron A /note="human cytomegalovirus intron A (Chapman et al., 1991)" regulatory 1604..1613 /regulatory_class="other" /note="vertebrate consensus sequence for strong initiation of translation (Kozak, 1987)" CDS 5366..5383 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 5409..5530 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5446..5465) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5500..5519 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5568..5585) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5739..6327) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5819..5838) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6498..7358) /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 7121..7140 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7359..7463) /gene="bla" /label=AmpR promoter primer_bind 7531..7549 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7587..7609) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7611..7631) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7616..7946 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7797..7932 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7859..7878 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 8083..8102 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(8169..8624) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8306..8327) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8518..8537 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8750..8772 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8764..8781 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8765..8781 /label=M13 fwd /note="common sequencing primer, one of multiple similar variants" ORIGIN 1 gtcgaggtct cgacggtatc gattgacatt gattattgac tagctagtta ttaatagtaa 61 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg 121 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg 181 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta 241 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtcc gccccctatt 301 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttacgggac 361 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt 421 tggcagtaca ccaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac 481 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt 541 cgtaataacc ccgccccgtt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat 601 ataagcagag ctcgtttagt gaaccgtcag atcgcctgga gacgccatcc acgctgtttt 661 gacctccata gaagacaccg ggaccgatcc agcctccgcg gccgggaacg gtgcattgga 721 acgcggattc cccgtgccaa gagtgacgta agtaccgcct atagactcta taggcacacc 781 cctttggctc ttatgcatgc tatactgttt ttggcttggg gcctatacac ccccgcttcc 841 atatgctata ggtgatggta tagcttagcc tataggtgtg ggttattgac cattattgac 901 cactccccta ttggtgacga tactttccat tactaatcca taacatggct ctttgccaca 961 actatctcta ttggctatat gccaatactc tgtccttcag agactgacac ggactctgta 1021 tttttacagg atggggtccc atttattatt tacaaattca catatacaac aacgccgtcc 1081 cccgtgcccg cagtttttat taaacatagc gtgggatctc cacgcgaatc tcgggtacgt 1141 gttccggaca tgggctcttc tccggtagcg gcggagcttc cacatccgag ccctggtccc 1201 atgcctccag cggctcatgg tcgctcggca gctccttgct cctaacagtg gaggccagac 1261 ttaggcacag cacaatgccc accaccacca gtgtgccgca caaggccgtg gcggtagggt 1321 atgtgtctga aaatgagctc ggagattggg ctcgcaccgc tgacgcagat ggaagactta 1381 aggcagcggc agaagaagat gcaggcagct gagttgttgt attctgataa gagtcagagg 1441 taactcccgt tgcggtgctg ttaacggtgg agggcagtgt agtctgagca gtactcgttg 1501 ctgccgcgcg cgccaccaga cataatagct gacagactaa cagactgttc ctttccatgg 1561 gtcttttctg cagaagcttg aattctcgag gatccgcggc cgcgccacca tggcaccggc 1621 cggacacatc ctcaccttgc tgctctgggg tcacctgctg gaactctgga ccccaggtca 1681 ctccgcgaac ccctcctacc ccaggctacg cctgtcacat aaagaacttt tggaactgaa 1741 taggacttca atatttcaaa gcccccttgg atttcttgat ctccatacaa tgctgctgga 1801 tgagtatcaa gaacggctct ttgtgggagg cagagacctt gtctattccc tgaacttgga 1861 acgagtcagt gacggctaca gagagatata ctggccgagc acagcagtaa aggtagaaga 1921 atgcataatg aaaggaaaag acgcaaatga gtgtgccaat tatatccggg ttttgcatca 1981 ctacaacagg acacaccttc tgacctgtgc tactggagct tttgatccac actgtgcctt 2041 catcagagtc gggcaccatt cagaggaacc cctgtttcac ctggagtcac acagatctga 2101 gagaggaagg ggcagatgtc cttttgaccc caactcctcc tttgtgtcca cgctagttgg 2161 gaatgagctg tttgctggac tctacagtga ctattggggc agagactcgg cgatcttccg 2221 cagcatgggg aagttaggcc atattcgcac tgagcatgac gatgagcggc tcctgaaaga 2281 accaaaattt gtaggttcat atatgattcc tgataacgaa gaccgagatg acaacaaaat 2341 gtactttttc tttactgaga aggcgctgga ggcggagaac aacgcccaca cgatctacac 2401 ccgagtgggg cggctgtgcg tgaatgacat gggaggacag agaatcctgg tgaacaagtg 2461 gagcactttc cttaaagcgc ggctggtttg ctcagtgccg ggaatgaatg gaatcgacac 2521 atactttgac gaactagagg atgtgttttt actgccgacc agagatccta agaatccagt 2581 gatatttgga ctgtttaata ctaccagcaa tatatttaga ggccatgctg tatgtgtgta 2641 tcacatgtca agtatccggg aagcctttaa tggcccatat gctcataaag aaggccctga 2701 ataccactgg tcactatatg aaggaaaagt cccctaccca aggcctggtt cctgtgccag 2761 caaagtaaac ggaggcaagt atggaaccac caaagattac cccgatgacg ccatccggtt 2821 cgcaaggatg catcctctaa tgtatcagcc cataaaacct gttcataaaa aaccaatact 2881 ggtaaaaaca gatggaaaat acaacctgag gcaacttgcc gtggatcggg tggaagcgga 2941 ggatggccag tatgacgtct tatttattgg gacagacaca ggaattgtgc tgaaagtaat 3001 cacaatttac aaccaagaaa cagagtggat ggaggaagtc attctagagg aacttcaaat 3061 attcaaggat ccagccccta tcatttctat ggaaatttct tcaaagagac aacagcttta 3121 cattggatca gcctctgctg tggcacaagt cagattccat cactgcgaca tgtatggcag 3181 tgcttgtgct gactgctgcc tggctcgaga cccgtactgt gcctgggatg gcatatcctg 3241 ctccaggtac tacccaacag gtgcacacgc aaagaggagg ttccgcaggc aggacgttcg 3301 gcatggcaac gccgcccaac agtgctttgg acagcaattt gttggagacg cgttggacag 3361 gactgaagag aggctggctt atggcataga gagcaacagt actctgttgg aatgcacccc 3421 gcgatcacta caagcaaaag tcatctggtt tgtacagaag ggacgcgacg taagaaaaga 3481 agaggtgaag acggatgaca gagttgtcaa gatggacttg ggcttgctct tcctcagagt 3541 acgcaagtca gatgcaggga cctatttttg ccagacagta gaacacaatt ttgtccatac 3601 tgtgcgtaaa atcaccttgg aggtggtcga agagcataaa gtggagggca tgtttcataa 3661 ggaccatgaa gaggaaagac atcacaagat gccctgccct cccttaagcg gtatgtctca 3721 ggggacaaaa ccgtggtaca aggaattctt gcagctgatt ggctacagca acttccagag 3781 agtggaagaa tactgcgaaa aggtgtggtg tacagataag aagaggaaaa agcttaaaat 3841 gtctccctcc aagtggaagt atgccaaccc ccaggaaggc gcgcctggat cttccggaat 3901 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg 3961 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg 4021 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga 4081 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa 4141 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg 4201 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg 4261 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa 4321 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc 4381 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga 4441 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg 4501 tgggggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta 4561 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg 4621 ccagggtgcc cggtacgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc 4681 tgtggcccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga 4741 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag 4801 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga 4861 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc 4921 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt 4981 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa 5041 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct 5101 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca 5161 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc 5221 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt 5281 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg 5341 caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta gagatcttat 5401 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa 5461 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca 5521 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac 5581 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata 5641 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa 5701 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct 5761 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa 5821 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg 5881 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca 5941 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa 6001 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg 6061 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg 6121 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga 6181 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc 6241 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag 6301 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac 6361 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc 6421 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag 6481 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt 6541 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag 6601 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca 6661 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact 6721 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca 6781 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg 6841 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc 6901 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg 6961 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca 7021 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt 7081 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc 7141 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc 7201 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca 7261 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa 7321 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat 7381 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa 7441 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa 7501 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc 7561 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg 7621 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc 7681 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta 7741 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc 7801 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta 7861 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct 7921 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc 7981 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg 8041 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg 8101 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc 8161 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat 8221 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata 8281 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt 8341 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc 8401 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa 8461 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg 8521 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt 8581 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg 8641 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg 8701 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga 8761 cgttgtaaaa cgacggccag tgaatt //