LOCUS       sequence-131232-        8435 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6D, isoform 6 protein,
            C-terminally fused to alkaline phosphatase + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8435)
  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 8435)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2015-12-3"; volume: "4. pii: e08149. doi"; pages: " 
            10.7554/eLife.08149"
COMMENT     Sequence Label: Sema6d.6-AP-His
FEATURES             Location/Qualifiers
     source          1..8435
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        25..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
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5015..5032
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5058..5179
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5095..5114)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5149..5168
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5217..5234)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5388..5976)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5468..5487)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6147..6938,6939..7007))
                     /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     6770..6789
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7008..7112)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7180..7198
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7236..7258)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7260..7280)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7265..7595
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7446..7581
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7508..7527
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7732..7751
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7818..8273)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7955..7976)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8167..8186
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8399..8421
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8413..8430
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8414..8430
                     /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 tggggttcct
     1621 tctgctttgg ttctgcgtgc tgttccttct ggtctccagg ttacgggcgg tcagcttccc
     1681 agaagacgat gagcccctca acacggttga ctatcactat tcaaggcaat atccggtttt
     1741 tagaggacgc ccttcaggca acgaatcgca gcacaggctg gactttcagc tgatgttgaa
     1801 aattcgagac acactttata ttgctggcag ggatcaagtc tatacagtga acttaaatga
     1861 aatcccccaa acagaggtga taccaagcaa gaagctgacg tggaggtcca gacagcagga
     1921 tcgagaaaat tgtgctatga aaggcaagca taaagatgaa tgccacaact tcatcaaagt
     1981 ctttgtccca agaaatgatg agatggtttt tgtctgtggt accaatgctt tcaacccgat
     2041 gtgcagatac tataggttga gaacgttaga gtatgatggg gaagaaatta gtggcctggc
     2101 acgatgcccg tttgatgccc gacaaaccaa tgtcgccctc tttgctgatg gaaaactcta
     2161 ttctgccaca gtggctgatt tcctggccag tgatgctgtc atttacagaa gcatgggaga
     2221 tggatctgcc cttcgcacaa taaaatacga ttccaagtgg atcaaagaac cacacttcct
     2281 tcatgccata gaatatggaa actatgtcta tttcttcttc agagaaatcg ccgtggaaca
     2341 taataactta ggcaaggctg tgtattcccg cgtggctcgc atttgtaaaa acgacatggg
     2401 tggctcacag cgggtcctgg agaaacactg gacttccttc cttaaggctc ggctgaactg
     2461 ctccgttcct ggagattcct ttttctactt cgacgtcctg cagtctataa cagacataat
     2521 ccaaatcaat ggcatcccca ctgtggttgg ggtcttcacc acacagctca acagcattcc
     2581 tggttctgca gtctgtgcct ttagcatgga cgacattgag aaagtgttca aagggcggtt
     2641 caaagagcag aaaaccccag actctgtttg gacagcagtt cccgaagaca aagtaccaaa
     2701 accaaggcct ggctgttgtg ccaaacacgg cctcgcagaa gcttacaaga cctccatcga
     2761 ctttccagat gacaccctgg ctttcatcaa gtcccacccg ctgatggact ctgccgtccc
     2821 acccattgcc gatgagccct ggttcacaaa gacacgggtc aggtacaggt tgacagccat
     2881 cgaagtggac cgttcagcag ggccatacca aaactacaca gtcatctttg ttggctctga
     2941 agctggcgtg gtacttaaag ttttggcaaa gaccagtcct ttctctctga atgacagtgt
     3001 attactcgaa gagattgaag cttataaccc agccaagtgc agcgccgaga gtgaggagga
     3061 cagaaaggtg gtctcattac agctggacaa ggatcaccat gctttatacg tggccttctc
     3121 tagctgcgtg gtccgcatcc ccctcagccg ctgtgagcgc tacggatcgt gtaaaaagtc
     3181 ttgcattgca tcacgtgacc cgtactgtgg ttggttaagc cagggagttt gtgagagagt
     3241 gaccctaggg atgctccctg gaggatatga gcaggacacg gagtacggca acacagccca
     3301 cctaggggac tgccacgaca tggaggtatc ctcatcttct gttaccactg tggcaagtag
     3361 cccagaaatt acatctaaag tgattgatac ctggagacct aaactgacga gctcccggaa
     3421 atttgtagtt caagatgacc caaatacttc tgattttact gatactatat caggtatccc
     3481 aaagggtgta cggtgggaag tccagtctgg agaatccaat cagatggtca ctagtggatc
     3541 ttccggaatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3601 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     3661 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     3721 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     3781 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     3841 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     3901 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     3961 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     4021 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     4081 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     4141 gatcctaggt ggaggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     4201 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     4261 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     4321 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     4381 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     4441 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4501 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4561 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4621 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     4681 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     4741 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     4801 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     4861 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     4921 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     4981 ccccgccggc accaccgacg ccgcgcaccc gggtcatcat caccatcacc attaatctag
     5041 agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5101 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5161 aactcatcaa tgtatcttat catgtctggt cgactctagc tagactcttc cgcttcctcg
     5221 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5281 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5341 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5401 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5461 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5521 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5581 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5641 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5701 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5761 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5821 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5881 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5941 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     6001 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     6061 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     6121 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6181 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6241 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6301 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6361 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6421 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6481 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6541 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6601 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6661 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6721 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6781 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6841 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6901 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6961 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     7021 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     7081 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     7141 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7201 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7261 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     7321 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     7381 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7441 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7501 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7561 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7621 ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     7681 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7741 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     7801 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7861 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7921 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7981 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     8041 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     8101 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     8161 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     8221 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     8281 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8341 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8401 cagtcacgac gttgtaaaac gacggccagt gaatt
//
