LOCUS       sequence-131233-        8474 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Sema6D, isoform x 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 8474)
  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 8474)
  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.x-AP-His
FEATURES             Location/Qualifiers
     source          1..8474
                     /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             5054..5071
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5097..5218
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5134..5153)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5188..5207
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5256..5273)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5427..6015)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5507..5526)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6186..6977,6978..7046))
                     /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     6809..6828
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7047..7151)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7219..7237
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7275..7297)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7299..7319)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7304..7634
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7485..7620
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7547..7566
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7771..7790
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7857..8312)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7994..8015)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8206..8225
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8438..8460
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8452..8469
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8453..8469
                     /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 gccacgacat
     3361 ggaggtatcc tcatcttctg ttaccactgt ggcaagtagc ccagaaatta catctaaagt
     3421 gattgatacc tggagaccta aactgacgag ctcccggaaa tttgtagttc aagatgaccc
     3481 aaatacttct gattttactg atactatatc aggtatccca aagggtgtac ggtgggaagt
     3541 ccagtctgga gaatccaatc agatggtcac tagtggatct tccggaatca tcccagttga
     3601 ggaggagaac ccggacttct ggaaccgcga ggcagccgag gccctgggtg ccgccaagaa
     3661 gctgcagcct gcacagacag ccgccaagaa cctcatcatc ttcctgggcg atgggatggg
     3721 ggtgtctacg gtgacagctg ccaggatcct aaaagggcag aagaaggaca aactggggcc
     3781 tgagataccc ctggccatgg accgcttccc atatgtggct ctgtccaaga catacaatgt
     3841 agacaaacat gtgccagaca gtggagccac agccacggcc tacctgtgcg gggtcaaggg
     3901 caacttccag accattggct tgagtgcagc cgcccgcttt aaccagtgca acacgacacg
     3961 cggcaacgag gtcatctccg tgatgaatcg ggccaagaaa gcagggaagt cagtgggagt
     4021 ggtaaccacc acacgagtgc agcacgcctc gccagccggc acctacgccc acacggtgaa
     4081 ccgcaactgg tactcggacg ccgacgtgcc tgcctcggcc cgccaggagg ggtgccagga
     4141 catcgctacg cagctcatct ccaacatgga cattgacgtg atcctaggtg gaggccgaaa
     4201 gtacatgttt cgcatgggaa ccccagaccc tgagtaccca gatgactaca gccaaggtgg
     4261 gaccaggctg gacgggaaga atctggtgca ggaatggctg gcgaagcgcc agggtgcccg
     4321 gtatgtgtgg aaccgcactg agctcatgca ggcttccctg gacccgtctg tgacccatct
     4381 catgggtctc tttgagcctg gagacatgaa atacgagatc caccgagact ccacactgga
     4441 cccctccctg atggagatga cagaggctgc cctgcgcctg ctgagcagga acccccgcgg
     4501 cttcttcctc ttcgtggagg gtggtcgcat cgaccatggt catcatgaaa gcagggctta
     4561 ccgggcactg actgagacga tcatgttcga cgacgccatt gagagggcgg gccagctcac
     4621 cagcgaggag gacacgctga gcctcgtcac tgccgaccac tcccacgtct tctccttcgg
     4681 aggctacccc ctgcgaggga gctccatctt cgggctggcc cctggcaagg cccgggacag
     4741 gaaggcctac acggtcctcc tatacggaaa cggtccaggc tatgtgctca aggacggcgc
     4801 ccggccggat gttaccgaga gcgagagcgg gagccccgag tatcggcagc agtcagcagt
     4861 gcccctggac gaagagaccc acgcaggcga ggacgtggcg gtgttcgcgc gcggcccgca
     4921 ggcgcacctg gttcacggcg tgcaggagca gaccttcata gcgcacgtca tggccttcgc
     4981 cgcctgcctg gagccctaca ccgcctgcga cctggcgccc cccgccggca ccaccgacgc
     5041 cgcgcacccg ggtcatcatc accatcacca ttaatctaga gatcttatta aagcagaact
     5101 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     5161 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     5221 atgtctggtc gactctagct agactcttcc gcttcctcgc tcactgactc gctgcgctcg
     5281 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     5341 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     5401 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     5461 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     5521 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     5581 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     5641 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     5701 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     5761 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     5821 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     5881 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     5941 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     6001 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     6061 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     6121 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     6181 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     6241 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     6301 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     6361 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     6421 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     6481 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     6541 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     6601 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     6661 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     6721 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     6781 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     6841 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     6901 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     6961 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     7021 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     7081 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     7141 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tctaagaaac cattattatc
     7201 atgacattaa cctataaaaa taggcgtatc acgaggccct ttcgtctcgc gcgtttcggt
     7261 gatgacggtg aaaacctctg acacatgcag ctcccggagg aatgtgtgtc agttagggtg
     7321 tggaaagtcc ccaggctccc caggcaggca gaagtatgca aagcatgcat ctcaattagt
     7381 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     7441 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     7501 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     7561 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     7621 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcgtcccg gagacggtca
     7681 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
     7741 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
     7801 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggaaat
     7861 tgtaagcgtt aatattttgt taaaattcgc gttaaatttt tgttaaatca gctcattttt
     7921 taaccaatag gccgaaatcg gcaaaatccc ttataaatca aaagaataga ccgagatagg
     7981 gttgagtgtt gttccagttt ggaacaagag tccactatta aagaacgtgg actccaacgt
     8041 caaagggcga aaaaccgtct atcagggcga tggcccacta cgtgaaccat caccctaatc
     8101 aagttttttg gggtcgaggt gccgtaaagc actaaatcgg aaccctaaag ggagcccccg
     8161 atttagagct tgacggggaa agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa
     8221 aggagcgggc gctagggcgc tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc
     8281 cgccgcgctt aatgcgccgc tacagggcgc gtccattcgc cattcaggct gcgcaactgt
     8341 tgggaagggc gatcggtgcg ggcctcttcg ctattacgcc agctggcgaa agggggatgt
     8401 gctgcaaggc gattaagttg ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg
     8461 acggccagtg aatt
//
