LOCUS       sequence-131228-        8306 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6D, isoform 1 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 8306)
  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 8306)
  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.1-AP-His
FEATURES             Location/Qualifiers
     source          1..8306
                     /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             4886..4903
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4929..5050
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4966..4985)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5020..5039
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5088..5105)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5259..5847)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5339..5358)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6018..6809,6810..6878))
                     /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     6641..6660
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6879..6983)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7051..7069
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7107..7129)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7131..7151)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7136..7466
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7317..7452
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7379..7398
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7603..7622
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7689..8144)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7826..7847)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8038..8057
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8270..8292
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8284..8301
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8285..8301
                     /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 atgctgctgt taaccgaaga cttctttgct ttccataacc acagccctgg
     3301 aggatatgag caggacacgg agtacggcaa cacagcccac ctaggggact gccacggtgt
     3361 acggtgggaa gtccagtctg gagaatccaa tcagatggtc actagtggat cttccggaat
     3421 catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg aggccctggg
     3481 tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca tcttcctggg
     3541 cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc agaagaagga
     3601 caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg ctctgtccaa
     3661 gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg cctacctgtg
     3721 cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct ttaaccagtg
     3781 caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga aagcagggaa
     3841 gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg gcacctacgc
     3901 ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg cccgccagga
     3961 ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg tgatcctagg
     4021 tggaggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc cagatgacta
     4081 cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc tggcgaagcg
     4141 ccagggtgcc cggtatgtgt ggaaccgcac tgagctcatg caggcttccc tggacccgtc
     4201 tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga tccaccgaga
     4261 ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc tgctgagcag
     4321 gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg gtcatcatga
     4381 aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca ttgagagggc
     4441 gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc actcccacgt
     4501 cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg cccctggcaa
     4561 ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag gctatgtgct
     4621 caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg agtatcggca
     4681 gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg cggtgttcgc
     4741 gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca tagcgcacgt
     4801 catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc cccccgccgg
     4861 caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta gagatcttat
     4921 taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa
     4981 atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca
     5041 atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc gctcactgac
     5101 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     5161 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     5221 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     5281 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     5341 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     5401 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     5461 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     5521 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     5581 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     5641 tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga
     5701 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     5761 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     5821 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     5881 gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc
     5941 ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag
     6001 taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt
     6061 ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag
     6121 ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca
     6181 gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact
     6241 ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca
     6301 gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg
     6361 tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc
     6421 atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg
     6481 gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca
     6541 tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt
     6601 atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc
     6661 agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc
     6721 ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca
     6781 tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa
     6841 aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat
     6901 tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa
     6961 aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtctaagaa
     7021 accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc ctttcgtctc
     7081 gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga ggaatgtgtg
     7141 tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg caaagcatgc
     7201 atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta
     7261 tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc
     7321 cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta
     7381 tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct
     7441 tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcgtcc
     7501 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     7561 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     7621 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     7681 gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt tttgttaaat
     7741 cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat caaaagaata
     7801 gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat taaagaacgt
     7861 ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac tacgtgaacc
     7921 atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc ggaaccctaa
     7981 agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga gaaaggaagg
     8041 gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca cgctgcgcgt
     8101 aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc gccattcagg
     8161 ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg ccagctggcg
     8221 aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc ccagtcacga
     8281 cgttgtaaaa cgacggccag tgaatt
//
