LOCUS Exported 10328 bp ds-DNA circular SYN 13-MAY-2021 DEFINITION Expresses the extracellular region of the PlexinD1 protein, C-terminally fused to alkaline phosphatase + 6X histidine tag.. ACCESSION . VERSION . KEYWORDS Plxnd1-AP-His SOURCE synthetic DNA construct ORGANISM synthetic DNA construct REFERENCE 1 (bases 1 to 10328) 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 10328) AUTHORS . TITLE Direct Submission JOURNAL Exported May 13, 2021 from SnapGene Server 1.1.58 http://www.snapgene.com FEATURES Location/Qualifiers source 1..10328 /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 6908..6925 /codon_start=1 /product="6xHis affinity tag" /label=6xHis /translation="HHHHHH" polyA_signal 6951..7072 /label=SV40 poly(A) signal /note="SV40 polyadenylation signal" primer_bind complement(6988..7007) /label=SV40pA-R /note="SV40 polyA, reverse primer" primer_bind 7042..7061 /label=EBV-rev /note="SV40 polyA terminator, reverse primer" primer_bind complement(7110..7127) /label=L4440 /note="L4440 vector, forward primer" rep_origin complement(7281..7869) /direction=LEFT /label=ori /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of replication" primer_bind complement(7361..7380) /label=pBR322ori-F /note="pBR322 origin, forward primer" CDS complement(8040..8900) /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 8663..8682 /label=Amp-R /note="Ampicillin resistance gene, reverse primer" promoter complement(8901..9005) /gene="bla" /label=AmpR promoter primer_bind 9073..9091 /label=pBRforEco /note="pBR322 vectors, upsteam of EcoRI site, forward primer" primer_bind complement(9129..9151) /label=pGEX 3' /note="pGEX vectors, reverse primer" primer_bind complement(9153..9173) /label=pBABE 3' /note="SV40 enhancer, reverse primer for pBABE vectors" promoter 9158..9488 /label=SV40 promoter /note="SV40 enhancer and early promoter" rep_origin 9339..9474 /label=SV40 ori /note="SV40 origin of replication" primer_bind 9401..9420 /label=SV40pro-F /note="SV40 promoter/origin, forward primer" primer_bind 9625..9644 /label=pRS-marker /note="pRS vectors, use to sequence yeast selectable marker" rep_origin complement(9711..10166) /direction=LEFT /label=f1 ori /note="f1 bacteriophage origin of replication; arrow indicates direction of (+) strand synthesis" primer_bind complement(9848..9869) /label=F1ori-F /note="F1 origin, forward primer" primer_bind 10060..10079 /label=F1ori-R /note="F1 origin, reverse primer" primer_bind 10292..10314 /label=M13/pUC Forward /note="In lacZ gene" primer_bind 10306..10323 /label=M13 Forward /note="In lacZ gene. Also called M13-F20 or M13 (-21) Forward" primer_bind 10307..10323 /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 tggctcgtcg 1621 cgccgcgggc ggcgcacccc ctagcgcccg ggccgccgcg gccgtcccct tgcgtccgcg 1681 ccctcactcg cggggccctg gtctgctgcc gctgcctctg ctgctgctgc tcggggcggc 1741 acgggccggc gccctagaga tccagcgccg tttcccctcg cccacgccca ccaacaactt 1801 cgccctggac ggcacggcgg gcaccgtgta cttggcggca gtgaaccgcc tgtaccaact 1861 gtcgagtgcc aacttgagcc tggaagccga ggcgaccgtg ggtcccgtgc cggacagccc 1921 gctgtgtcac gccccgcagc tcccgcaggc ctcgtgcgag cacccgcggc gcctcacgga 1981 caactacaac aaaatcctgc agttggaccc gggccagggt ctggtggtcg cgtgcggctc 2041 catctaccag ggtctgtgcc agctgaggcg ccggggcaac atctcagccc tggccgtgag 2101 ctttccgcct gccgcgccga ccgcagaacc ggtcaccgtg ttccccagca tgctcaacgt 2161 ggccgccaac caccccaacg cgtccactgt gggactggtg ctgccgccta cctcgggcac 2221 cgggggcagc cgtctgctcg tgggcgccac gtacaccggc ttcggcagcg ctttcttccc 2281 gcgcaaccgt agcctagaag accaccgctt cgagaacacg cccgagatcg ctatccgctc 2341 cctggacgcg cgtggagact tggccaagct cttcaccttc gaccttaacc cgtcggacga 2401 taacatcctg aagatcaagc agggcgccaa ggagcagcac aagctgggct tcgtgcgtgc 2461 cttcttgcac ccggcggtgc caccgcacag cgcgcagccc tacgcgtacc tggcgctcaa 2521 cagcgaggcg cgtgcgggcg acaaggacag ccaggcgcgc agcctgctgg cgcgcatctg 2581 cctgccccgc ggcgcgggtg gcgacgccaa gaagctcacc gagtcctaca tccaactggg 2641 cttgcagtgc gcgggcggcg cgggccgcgg cgacctctac agccgcctcg tgtcggtttt 2701 ccccgcgcgc gagcagttct tcgctgtctt cgagaggccc cagggcgccc caggtgcccg 2761 caacgccccg gccgcgcttt gcgccttccg cttcgacgac gtgcaggctg ccattcgtgc 2821 agcgcgcacc gcctgcttcg tggagccggc gcccgacgtg gtggcggtgt tggacagtgt 2881 ggtgcagggc accgggccgg cctgcgagag caagcgcaac atacagctgc agccggagca 2941 actggattgc ggagcggccc acctgcagca cccactgacc atcctgcagc cgctgagggc 3001 atcgcccgtg ttccgtgctc cagggctcac ggccgtggct gtggccagtg ccaacaacta 3061 cacggccgtc tttctgggca ccgccacagg gaggctcctc aagatcagcc tgaacgagag 3121 catgcaggta gtaagcaggc gagtgctgac tgtagcctat ggggagcctg tgcatcacgt 3181 catgcagttt gaccccatgg atcctggtta cctatacctg atgacatccc accagatggc 3241 ccgagtgaag gtggcagcgt gtgaggtaca ctccacctgc ggggactgcg tgggtgcggc 3301 cgatgcctac tgtggttggt gcactctgga gacccggtgc acactccagc aggattgcac 3361 caactccagc cagccacatt tctggaccag tgccagtgag ggccccagcc gctgccctgc 3421 catgacagta ctgccctcgg agattgatgt gcaccgggac tacacaggta tgatcttaca 3481 gatctcagga agcctgccca gcctcagcgg catggagatg gcttgtgact atggaaatgg 3541 cgttcgaacg gtggcccggg tacctggccc tgcctatgat catcagattg cctactgcaa 3601 tctcctgccc agggcccagt ttccatcctt tcctgctggc caggaccacg tgaccgttga 3661 gatgtctgta agggtcaaag gacacaacat tgtctcagcc aatttcacca tctacgactg 3721 cagccgaatt ggacaagtct acccccatac agcctgtacc agctgcctgt ccacacagtg 3781 gccttgctcc tggtgcatcc agctgcattc atgtgtctcc aaccagtctc agtgccagga 3841 ctcgccaaac cccacgagtc ctcaggactg tccccagatc ctgccctcgc ccctagcgcc 3901 cgtgcccaca ggtggctccc aagacatcct ggtgcccctg actaaagcca ccttcttcca 3961 tggttcctcc ctcgagtgca gctttgggct ggaagagagc tttgaggctg tatgggcgaa 4021 taactcactg gtccgctgca accaagtggt gctgcacaca acccagaaga gccaggtatt 4081 tccactgagt ctgaagctga aggggccgcc agaccgattc ctagacagcc ctaaccccat 4141 gacagttgtg gtctacaact gtgctatggg cagccctgac tgttcccagt gcctgggccg 4201 tgaggacctg ggtcacctct gtgtttggaa tgatggctgt cgtctaagag ggcccctgca 4261 gccactccct ggcacctgcc cagcccctga aatccgagcg attgagcctc tgagtggccc 4321 cttggacggt gggactttgc tgaccatccg tggcaggaac ttgggccgtc ggctcagtga 4381 tgtggcacat ggtgtgtgga ttggcagtgt ggcctgtgaa cccctggctg acagatacac 4441 cgtttcagag gagatcgtgt gtgccacagg gcctgccgca ggggccttct cagacgtggt 4501 aacggtgaac gtctccaaag aaggcaggtc tcgggaacag ttctcctatg tgctgcccac 4561 agtccactca ctggagcctt ccatgggccc aaaggccggg ggtacaagga tcaccattca 4621 cggcagtgac ctcaacgtgg gctctatgct ccaggtcctg gtgaatgaca cggacccctg 4681 cacagatctt acgcgcacag ccaccagcat cacctgcact gtgccagggg gtaccctgcc 4741 ctctccagtg cctgtgtgtg tgcgcttcga gagccggggc tgcgtgcacg gaaacctcac 4801 cttctggtac atgcagaacc cagtcatcac agccatcagc ccaggccgca gccctgtcag 4861 tggcggcagg accatcactg tggctggcga acgcttccac atggtgcaga atgtatcaat 4921 ggctgtacac cacattggcc gggagcccac gttctgcaag gttctcaact ccacactcat 4981 cacctgccca tctcctggag ccctgagcaa tgcttcggcg cctgtagact tcttcatcaa 5041 tggccgggca tatgcagacg aggcagccga ggagctgctg gaccctgcag aggcacagag 5101 gggcagccgg ttccgcctag actacctccc caacccacag ttctccacag ccaagaggga 5161 gaagtggatc aaacatcacc caggagagcc gctcaccctc gtcatccata aggagcaaga 5221 cagcctgggg ctggagagcc atgagtacca catcaagatt ggccaggtgt cctgcgacat 5281 ccaaatcatc tcagacagag tcatccactg ctcagtcaat gagtcgctgg gcacggctga 5341 aggacagctg cccatcacaa tccaggtggg gaacttcaac cagaccatcg ccacactgca 5401 actggggggc agcgagacgg gcgcgcctgg atcttccgga atcatcccag ttgaggagga 5461 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca 5521 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc 5581 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat 5641 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa 5701 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt 5761 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa 5821 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac 5881 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa 5941 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc 6001 tacgcagctc atctccaaca tggacattga cgtgatccta ggtgggggcc gaaagtacat 6061 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag 6121 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt 6181 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg 6241 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc 6301 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt 6361 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc 6421 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga 6481 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta 6541 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc 6601 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc 6661 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct 6721 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca 6781 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg 6841 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca 6901 cccgggtcat catcaccatc accattaatc tagagatctt attaaagcag aacttgttta 6961 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat 7021 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct 7081 ggtcgactct agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt 7141 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca 7201 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa 7261 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat 7321 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc 7381 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc 7441 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt 7501 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac 7561 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg 7621 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca 7681 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc 7741 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa 7801 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa 7861 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac 7921 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta 7981 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt 8041 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata 8101 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc 8161 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac 8221 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag 8281 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac 8341 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc 8401 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg 8461 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc 8521 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct 8581 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc 8641 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc 8701 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc 8761 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc 8821 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca 8881 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt 8941 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt 9001 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca 9061 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac 9121 ggtgaaaacc tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa 9181 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa 9241 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca 9301 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca 9361 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg 9421 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct 9481 tttgcaaaaa gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt 9541 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg 9601 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata 9661 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag 9721 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca 9781 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag 9841 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg 9901 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt 9961 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag 10021 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc 10081 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc 10141 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa 10201 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca 10261 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc 10321 agtgaatt //