LOCUS Exported 8327 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the extracellular region of the Sema6C protein, C-terminally fused to alkaline phosphatase + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Sema6c-AP-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8327) 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 8327) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8327 /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)" CDS 4907..4924 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 4950..5071 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4987..5006) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5041..5060 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5109..5126) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5280..5868) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5360..5379) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6039..6899) /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 6662..6681 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6900..7004) /gene="bla" /label=AmpR promoter primer_bind 7072..7090 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7128..7150) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7152..7172) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7157..7487 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7338..7473 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7400..7419 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 7624..7643 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(7710..8165) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7847..7868) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8059..8078 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8291..8313 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8305..8322 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8306..8322 /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 tgccgcgtgc 1621 cccccactcc atgcccctgc tgctcctgct gctgttgctg tcatcgctcc cccaagccca 1681 ggctgccttt ccccaggacc ccacccctct gttgacctct gaccttcaag gtgcctctcc 1741 atcatcctgg ttccggggcc tggaggacga tgctgtggct gcggaacttg ggctggactt 1801 tcagagattc ctgaccttga accggacctt gcttgtggct gcccgggatc acgttttctc 1861 cttcgatctt caagcccaag aagaagggga ggggctggtg cccaacaagt ttctgacatg 1921 gaggagtcaa gatatggaga actgtgcagt ccggggaaag ctgacggacg aatgctacaa 1981 ctacatccgt gttcttgttc cctggaactc gcagacactt cttgcctgtg gaacaaattc 2041 cttcagccct atgtgccgca gctatgggat aacatctctg caacaggagg gtgaggagct 2101 gagtgggcag gctcgatgcc cctttgatgc cacccagtcc actgtggcca tctttgcaga 2161 gggcagtttg tactcagcca cagcagcaga tttccaggcc agtgatgctg tagtttacag 2221 aagtcttgga cctcagcctc cactccgttc tgcaaagtat gactccaagt ggcttcgaga 2281 gccacacttt gtctatgcgt tggagcatgg agagcacgtc tacttcttct tccgagaagt 2341 ctctgtggag gatgcccgct tagggagggt gcagttttcc agggtagccc gggtatgtaa 2401 acgtgacatg ggtggctcac ctcgggcctt ggatcgccac tggacatcct tccttaagct 2461 gaggctcaac tgctctgtcc ctggggactc taccttctac tttgacgtct tacagtcctt 2521 aactgggcct gtgaacctgc atggccgctc cgctctcttt ggggtcttca ctactcagac 2581 caatagcatt cctgggtctg cagtctgcgc cttctaccta gatgatattg aacgtggctt 2641 tgagggcaag ttcaaggagc agaggagtct ggatggggcc tggactcctg tgtctgagga 2701 caaggtcccc tcacccaggc cagggtcctg tgcaggtgtg ggtgcagctg cctcgttctc 2761 ctcctctcaa gacctacctg atgatgtcct gctcttcatc aaggcacacc cgctgctgga 2821 tcctgctgtg ccacccgcca cccatcaacc gctcctcact ctgactagca gggccctgct 2881 gacccaggta gctgtggacg gaatggctgg tccccacaga aatactactg tcctgtttct 2941 tggctccaac gatgggacag tgctgaaggt gctacctcca gggggtcagt ctttgggctc 3001 tgagcctatc gtcttggaag agattgatgc ctacagccat gcccggtgca gtgggaagcg 3061 gtcaccccga gctgcgcggc ggatcatagg gctggagctg gacactgagg gtcacaggct 3121 ttttgtggcc tttcctggat gtatcgtcta cctctctctc agccggtgtg cccggcatgg 3181 agcatgtcag aggagctgcc tggcttctct ggacccatac tgtggatggc atcgatccag 3241 aggctgcatg agtatcaggg gacctggtgg gactgatgtc gatctgactg ggaaccagga 3301 atccacagag cacggtgact gccaagatgg agcgaccggg agtcagtctg gccctggaga 3361 ttctgcttat ggcgtgcgca gggacctttc cccagcctca gcctcccgat ccattcccgg 3421 cgcgcctgga tcttccggaa tcatcccagt tgaggaggag aacccggact tctggaaccg 3481 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa 3541 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat 3601 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt 3661 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc 3721 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc 3781 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa 3841 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc 3901 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt 3961 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat 4021 ggacattgac gtgatcctag gtgggggccg aaagtacatg tttcgcatgg gaaccccaga 4081 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt 4141 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat 4201 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat 4261 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc 4321 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg 4381 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt 4441 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt 4501 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat 4561 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg 4621 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag 4681 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg 4741 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga 4801 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg 4861 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggtcatc atcaccatca 4921 ccattaatct agagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca 4981 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt 5041 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gtcgactcta gctagactct 5101 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca 5161 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac 5221 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt 5281 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg 5341 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc 5401 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc 5461 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc 5521 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac 5581 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt 5641 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct 5701 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc 5761 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt 5821 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg 5881 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc 5941 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa 6001 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag 6061 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg 6121 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga 6181 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag 6241 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa 6301 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc 6361 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca 6421 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg 6481 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat 6541 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc 6601 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg 6661 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg 6721 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt 6781 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca 6841 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata 6901 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac 6961 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa 7021 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt 7081 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg 7141 cagctcccgg aggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag 7201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag 7261 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc 7321 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc 7381 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat 7441 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag 7501 cttgtatatc cattttcgtc ccggagacgg tcacagcttg tctgtaagcg gatgccggga 7561 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact 7621 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca 7681 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt 7741 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat 7801 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa 7861 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg 7921 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa 7981 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc 8041 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag 8101 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg 8161 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct 8221 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg 8281 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaatt //