LOCUS Exported 8750 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses the Sema3F protein (truncated at cleavage site P3; ie, long and contains a 93 nt deletion in exon 3), C-terminally fused to alkaline phosphatase + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Sema3f(L,-)-AP-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 8750) 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 8750) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..8750 /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 5330..5347 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 5373..5494 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(5410..5429) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 5464..5483 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(5532..5549) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(5703..6291) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(5783..5802) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(6462..7322) /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 7085..7104 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(7323..7427) /gene="bla" /label=AmpR promoter primer_bind 7495..7513 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(7551..7573) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(7575..7595) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 7580..7910 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 7761..7896 /label=SV40 ori /note="SV40 origin of replication" primer_bind 7823..7842 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 8047..8066 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(8133..8588) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(8270..8291) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 8482..8501 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 8714..8736 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 8728..8745 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 8729..8745 /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 tgcttgtcac 1621 tgccttcatc ctctgggctt ccctgctgac cggggcctgg cctgctaccc ctatccagga 1681 ccagcttcca gccacaccta gagtccgact gtcattcaaa gaacttaagg ccaccggtac 1741 cgcccacttc ttcaactttc tgctcaacac tacagactac agaatcctgc tcaaggatga 1801 ggaccatgac cgcatgtatg tgggcagcaa ggactacgtg ctgtccctgg acctgcatga 1861 catcaaccga gagcccctta ttatccattg ggcagcctcc ccgcagcgca ttgaggagtg 1921 catactgtca ggcaaggatg gcaatggaga gtgtggtaac ttcgtccggc tcatccagcc 1981 ttggaaccga acacacctgt atgtgtgtgg gaccggtgcc tacaacccca tgtgcaccta 2041 tgtgaaccgt ggccgtcgcg cacaggatta catcttctac ctggagcctg agaaactgga 2101 gtcaggaaaa ggcaaatgtc cctatgaccc caagctggac acagcctcag ccctcatcaa 2161 tgaggaactc tatgcaggag tgtacatcga ttttatgggt accgatgcag ccattttccg 2221 cacgctgggg aagcagacag ccatgcgcac agatcagtac aactcccggt ggctcaatga 2281 tccttccttc atacatgctg agctcatccc tgacagcgca gagcgcaatg acgataaact 2341 ctacttcttc ttccgagagc gctcggcaga ggcgccacag aaccctgctg tgtatgcccg 2401 catcgggcgc atctgcctca acgatgacgg tggccattgc tgcctggtca acaagtggag 2461 cacattcctg aaggctcggc tcgtctgctc cgtgccaggc gaggatggta tcgagaccca 2521 cttcgacgaa ctccaggatg tgtttgtcca gcagacccag gacatcagga acccagtcat 2581 ttatgctgtc tttacctctt caggctctgt gttccgaggg tctgcagtgt gtgtctactc 2641 catggctgat atccgcatgg tcttcaatgg gccttttgct cacaaggagg gccccaacta 2701 tcaatggatg cccttctcag gaaagatgcc ctatcctcgg cctggcacat gccctggtgg 2761 aaccttcacg ccctcgatga agtccactaa ggactaccct gatgaagtga tcaacttcat 2821 gcgcactcat ccgctcatgt accaggccgt ctacccgctc cagcggcgac ccctggtggt 2881 ccgtacaggc gctccctacc gcctcaccac tgtcgctgtg gaccaggtgg atgcagctga 2941 tggacgctat gaggtgcttt tcctgggcac agaccgcggg acagtgcaga aggtcatcgt 3001 gctgcccaag gacgaccagg aggtggaaga gctgatgcta gaggaagtgg aggtcttcaa 3061 ggaaccagca cctgttaaga ccatgaccat ctcttccaag aggcaacaac tgtatgtggc 3121 ctcagccgtg ggtgtcacac acctgagcct gcaccgctgc caggcctatg gagctgcctg 3181 tgccgactgc tgcctcgccc gcgatcccta ctgtgcctgg gatggccagg cctgctctcg 3241 ctacacagca tcctccaaga ggcggagtcg ccggcaagat gtccgtcacg ggaaccctat 3301 cagacagtgc cgtggtttca attccaatgc aaacaagaat gccgtagagt ctgtgcagta 3361 cggagtggcc ggcagcgcag ctttccttga gtgtcagccc cgctcgcctc aagccactgt 3421 caagtggctg ttccagcgag atcccagtga tcggcgccgt gagattcgcg cagaggaccg 3481 cttcctgcgt acagagcagg ggctgttgct tcgcgccctg cagcttggtg accgcggcct 3541 ctactcctgc acggccactg agaacaactt caagcacatc gtcacgcggg tgcagctgca 3601 tgtactgggc cgggacgccg tccatgctgc cctcttcccc ccactggctg tgagcgtccc 3661 tccacctcct ggcacaggcc ctccaacgcc tccttaccag gagctggccc agctcctagc 3721 ccagccggaa gtgggcctca tccatcagta ctgccagggt tactggcgcc acgtgccccc 3781 cagccccagg gaggcgccag gagcacttag gcctcctgaa ctccaggacc agaaaaagcc 3841 gggcgcgcct ggatcttccg gaatcatccc agttgaggag gagaacccgg acttctggaa 3901 ccgcgaggca gccgaggccc tgggtgccgc caagaagctg cagcctgcac agacagccgc 3961 caagaacctc atcatcttcc tgggcgatgg gatgggggtg tctacggtga cagctgccag 4021 gatcctaaaa gggcagaaga aggacaaact ggggcctgag atacccctgg ccatggaccg 4081 cttcccatat gtggctctgt ccaagacata caatgtagac aaacatgtgc cagacagtgg 4141 agccacagcc acggcctacc tgtgcggggt caagggcaac ttccagacca ttggcttgag 4201 tgcagccgcc cgctttaacc agtgcaacac gacacgcggc aacgaggtca tctccgtgat 4261 gaatcgggcc aagaaagcag ggaagtcagt gggagtggta accaccacac gagtgcagca 4321 cgcctcgcca gccggcacct acgcccacac ggtgaaccgc aactggtact cggacgccga 4381 cgtgcctgcc tcggcccgcc aggaggggtg ccaggacatc gctacgcagc tcatctccaa 4441 catggacatt gacgtgatcc taggtggggg ccgaaagtac atgtttcgca tgggaacccc 4501 agaccctgag tacccagatg actacagcca aggtgggacc aggctggacg ggaagaatct 4561 ggtgcaggaa tggctggcga agcgccaggg tgcccggtac gtgtggaacc gcactgagct 4621 catgcaggct tccctggacc cgtctgtggc ccatctcatg ggtctctttg agcctggaga 4681 catgaaatac gagatccacc gagactccac actggacccc tccctgatgg agatgacaga 4741 ggctgccctg cgcctgctga gcaggaaccc ccgcggcttc ttcctcttcg tggagggtgg 4801 tcgcatcgac catggtcatc atgaaagcag ggcttaccgg gcactgactg agacgatcat 4861 gttcgacgac gccattgaga gggcgggcca gctcaccagc gaggaggaca cgctgagcct 4921 cgtcactgcc gaccactccc acgtcttctc cttcggaggc taccccctgc gagggagctc 4981 catcttcggg ctggcccctg gcaaggcccg ggacaggaag gcctacacgg tcctcctata 5041 cggaaacggt ccaggctatg tgctcaagga cggcgcccgg ccggatgtta ccgagagcga 5101 gagcgggagc cccgagtatc ggcagcagtc agcagtgccc ctggacgaag agacccacgc 5161 aggcgaggac gtggcggtgt tcgcgcgcgg cccgcaggcg cacctggttc acggcgtgca 5221 ggagcagacc ttcatagcgc acgtcatggc cttcgccgcc tgcctggagc cctacaccgc 5281 ctgcgacctg gcgccccccg ccggcaccac cgacgccgcg cacccgggtc atcatcacca 5341 tcaccattaa tctagagatc ttattaaagc agaacttgtt tattgcagct tataatggtt 5401 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta 5461 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact ctagctagac 5521 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta 5581 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag 5641 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg 5701 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg 5761 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg 5821 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga 5881 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc 5941 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt 6001 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact 6061 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg 6121 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt 6181 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt 6241 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct 6301 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg 6361 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt 6421 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt 6481 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc 6541 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg 6601 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc 6661 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg 6721 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca 6781 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga 6841 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct 6901 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg 6961 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca 7021 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata 7081 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct 7141 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact 7201 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa 7261 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc 7321 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga 7381 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga 7441 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg 7501 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac 7561 atgcagctcc cggaggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg 7621 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc 7681 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag 7741 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc 7801 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc 7861 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg 7921 gagcttgtat atccattttc gtcccggaga cggtcacagc ttgtctgtaa gcggatgccg 7981 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta 8041 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc 8101 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa 8161 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa 8221 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa 8281 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca 8341 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg 8401 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc 8461 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc 8521 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca 8581 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc 8641 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta 8701 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt //