LOCUS Exported 7763 bp ds-DNA circular SYN 12-MAY-2021 DEFINITION Expresses the extracellular region of the Sema4A protein, C-terminally fused to the Fc region of human IgG1 + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Sema4a-Fc-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 7763) 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 7763) AUTHORS . TITLE Direct Submission JOURNAL Exported May 12, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..7763 /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 4343..4360 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 4386..4507 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(4423..4442) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 4477..4496 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(4545..4562) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(4716..5304) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(4796..4815) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(5475..6335) /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 6098..6117 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(6336..6440) /gene="bla" /label=AmpR promoter primer_bind 6508..6526 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(6564..6586) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(6588..6608) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 6593..6923 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 6774..6909 /label=SV40 ori /note="SV40 origin of replication" primer_bind 6836..6855 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 7060..7079 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(7146..7601) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(7283..7304) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 7495..7514 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 7727..7749 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 7741..7758 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 7742..7758 /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 tggccctacc 1621 atccctgggc caggactcat ggagtctcct gcgtgttttt ttcttccaac tcttcctgct 1681 gccatcactg ccacctgctt ctgggactgg tggtcagggg cccatgccca gagtcaaata 1741 ccatgctgga gacgggcaca gggccctcag cttcttccaa caaaaaggcc tccgagactt 1801 tgacacgctg ctcctgagtg acgatggcaa cactctctat gtgggggctc gagaggccgt 1861 cctggccttg aatatccaga acccaggaat cccaaggcta aagaacatga taccctggcc 1921 agccagtgag agaaaaaaga ccgaatgtgc ctttaagaag aagagcaatg agacacagtg 1981 tttcaacttc attcgagtcc tggtctctta caatgctact cacctctatg cctgtgggac 2041 ctttgccttc agccctgcct gtaccttcat tgaactccaa gattccctcc tgttgcccat 2101 cttgatagac aaggtcatgg acgggaaggg ccaaagcccc tttgaccctg ttcacaagca 2161 cacagctgtc ttggtcgatg ggatgcttta ttccggcacc atgaacaact tcctgggcag 2221 cgagcccatc ctgatgcgga cactgggatc ccagcctgtt ctcaagactg acatcttctt 2281 acgctggctg cacgcggatg cctccttcgt ggcagccatt ccatccaccc aggtcgtcta 2341 tttcttcttt gaggagacag ccagcgagtt tgacttcttt gaagagctgt atatatccag 2401 ggtggctcaa gtctgcaaga acgacgtggg cggtgaaaag ctgctgcaga agaagtggac 2461 caccttcctc aaagcccagt tgctctgcgc tcagccaggg cagctgccat tcaacatcat 2521 ccgccacgcg gtcctgctgc ccgccgattc tccctctgtt tcccgcatct acgcagtctt 2581 tacctcccag tggcaggttg gcgggaccag gagctcagca gtctgtgcct tctctctcac 2641 ggacattgag cgagtcttta aagggaagta caaggagctg aacaaggaga cctcccgctg 2701 gaccacttac cggggctcag aggtcagccc gaggccaggc agttgctcca tgggcccctc 2761 ctctgacaaa gccttgacct tcatgaagga ccattttctg atggatgagc acgtggtagg 2821 aacacccctg ctggtgaagt ctggtgtgga gtacacacgg cttgctgtgg agtcagctcg 2881 gggccttgat gggagcagcc atgtggtcat gtatctgggt acctccacgg ggtccctgca 2941 caaggctgtg gtgcctcagg acagcagtgc ttatctcgtg gaggagattc agctgagccc 3001 tgactctgag cctgttcgaa acctgcagct ggcccccgcc cagggtgcag tgtttgcagg 3061 cttctctgga ggcatctgga gagttcccag ggccaattgc agtgtctacg agagctgtgt 3121 ggactgtgtg cttgccaggg accctcactg tgcctgggac cctgaatcaa gactctgcag 3181 ccttctgtct ggctctacca agccttggaa gcaggacatg gaacgcggca acccggagtg 3241 ggtatgcacc cgtggcccca tggccaggag cccccggcgt cagagccccc ctcaactaat 3301 taaagaagtc ctgacagtcc ccaactccat cctggagctg ccctgccccc acctgtcagc 3361 actggcctct taccactgga gtcatggcca agccaaaatc tcagaagcct ctgctaccgt 3421 ctacaatggc tccctcttgc tgctgccgca ggatggtgtc gggggcctct accagtgtgt 3481 ggcgactgag aacggctact cataccctgt ggtctcctat tgggtagaca gccaggacca 3541 gcccctggcg ctggaccctg agctggcggg cgttccccgt gagcgtgtgc aggtcccgct 3601 gaccagggtc ggaggcggag cttccatggc tgcccagcgg tcctacggcg cgcctgacaa 3661 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct 3721 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt 3781 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt 3841 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt 3901 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa 3961 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca 4021 gccccgagaa ccacaggtgt ataccctgcc cccatcccgg gatgagctga ccaagaacca 4081 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga 4141 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg 4201 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt 4261 cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc 4321 cctgtctccg ggtaaaggcg gccatcatca ccatcaccat taatctagag atcttattaa 4381 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt 4441 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg 4501 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg 4561 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg 4621 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag 4681 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac 4741 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga 4801 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt 4861 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc 4921 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc 4981 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta 5041 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat 5101 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca 5161 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct 5221 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt 5281 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct 5341 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc 5401 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa 5461 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta 5521 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc 5581 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat 5641 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta 5701 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt 5761 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt 5821 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg 5881 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc 5941 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc 6001 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg 6061 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga 6121 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta 6181 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct 6241 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag 6301 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga 6361 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat 6421 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc 6481 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg 6541 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca 6601 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc 6661 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc 6721 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc 6781 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt 6841 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt 6901 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg 6961 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt 7021 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac 7081 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca 7141 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag 7201 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac 7261 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga 7321 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc 7381 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg 7441 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa 7501 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac 7561 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg 7621 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa 7681 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt 7741 tgtaaaacga cggccagtga att //