LOCUS Exported 7994 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the Sema3E protein (truncated at cleavage site P3; ie, long), C-terminally fused to the Fc region of human IgG1 + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Sema3e(L)-Fc-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7994) 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 7994) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7994 /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 4574..4591 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 4617..4738 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4654..4673) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4708..4727 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4776..4793) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4947..5535) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5027..5046) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5706..6566) /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 6329..6348 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6567..6671) /gene="bla" /label=AmpR promoter primer_bind 6739..6757 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6795..6817) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6819..6839) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6824..7154 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7005..7140 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7067..7086 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 7291..7310 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(7377..7832) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7514..7535) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7726..7745 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7958..7980 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7972..7989 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7973..7989 /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 gcgcctgaca aaactcacac 3901 atgcccaccg tgcccagcac ctgaactcct ggggggaccg tcagtcttcc tcttcccccc 3961 aaaacccaag gacaccctca tgatctcccg gacccctgag gtcacatgcg tggtggtgga 4021 cgtgagccac gaagaccctg aggtcaagtt caactggtac gtggacggcg tggaggtgca 4081 taatgccaag acaaagccgc gggaggagca gtacaacagc acgtaccgtg tggtcagcgt 4141 cctcaccgtc ctgcaccagg actggctgaa tggcaaggag tacaagtgca aggtctccaa 4201 caaagccctc ccagccccca tcgagaaaac catctccaaa gccaaagggc agccccgaga 4261 accacaggtg tataccctgc ccccatcccg ggatgagctg accaagaacc aggtcagcct 4321 gacctgcctg gtcaaaggct tctatcccag cgacatcgcc gtggagtggg agagcaatgg 4381 gcagccggag aacaactaca agaccacgcc tcccgtgctg gactccgacg gctccttctt 4441 cctctacagc aagctcaccg tggacaagag caggtggcag caggggaacg tcttctcatg 4501 ctccgtgatg catgaggctc tgcacaacca ctacacgcag aagagcctct ccctgtctcc 4561 gggtaaaggc ggccatcatc accatcacca ttaatctaga gatcttatta aagcagaact 4621 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata 4681 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc 4741 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg 4801 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca 4861 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac 4921 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac 4981 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg 5041 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac 5101 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat 5161 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag 5221 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac 5281 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt 5341 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt 5401 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc 5461 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga 5521 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac 5581 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc 5641 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct 5701 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca 5761 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct 5821 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca 5881 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc 5941 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg 6001 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct 6061 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa 6121 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta 6181 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc 6241 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg 6301 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa 6361 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg 6421 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc 6481 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg 6541 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat 6601 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata 6661 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc 6721 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt 6781 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg 6841 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt 6901 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc 6961 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc 7021 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg 7081 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc 7141 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca 7201 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg 7261 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc 7321 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat 7381 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt 7441 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg 7501 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt 7561 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc 7621 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg 7681 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa 7741 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc 7801 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt 7861 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt 7921 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg 7981 acggccagtg aatt //