LOCUS       sequence-131229-        8267 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Expresses the extracellular region of the Sema6D, isoform 2 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 8267)
  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 8267)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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.2-AP-His
FEATURES             Location/Qualifiers
     source          1..8267
                     /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             4847..4864
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4890..5011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4927..4946)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4981..5000
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5049..5066)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5220..5808)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5300..5319)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5979..6770,6771..6839))
                     /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     6602..6621
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6840..6944)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7012..7030
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7068..7090)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7092..7112)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7097..7427
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7278..7413
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7340..7359
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7564..7583
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7650..8105)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7787..7808)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7999..8018
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8231..8253
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8245..8262
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8246..8262
                     /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 ggcagcagga
     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 tgccacggtg tacggtggga agtccagtct ggagaatcca atcagatggt
     3361 cactagtgga tcttccggaa tcatcccagt tgaggaggag aacccggact tctggaaccg
     3421 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     3481 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     3541 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     3601 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     3661 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     3721 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     3781 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     3841 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     3901 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     3961 ggacattgac gtgatcctag gtggaggccg aaagtacatg tttcgcatgg gaaccccaga
     4021 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     4081 gcaggaatgg ctggcgaagc gccagggtgc ccggtatgtg tggaaccgca ctgagctcat
     4141 gcaggcttcc ctggacccgt ctgtgaccca tctcatgggt ctctttgagc ctggagacat
     4201 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     4261 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     4321 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     4381 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     4441 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     4501 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     4561 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     4621 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     4681 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     4741 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     4801 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggtcatc atcaccatca
     4861 ccattaatct agagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca
     4921 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4981 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gtcgactcta gctagactct
     5041 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     5101 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     5161 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     5221 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     5281 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     5341 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     5401 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     5461 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     5521 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     5581 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5641 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5701 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5761 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5821 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5881 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5941 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     6001 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     6061 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     6121 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     6181 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     6241 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     6301 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     6361 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     6421 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     6481 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     6541 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6601 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6661 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6721 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6781 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6841 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6901 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6961 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     7021 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     7081 cagctcccgg aggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag
     7141 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     7201 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     7261 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     7321 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     7381 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     7441 cttgtatatc cattttcgtc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     7501 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     7561 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     7621 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     7681 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7741 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7801 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7861 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7921 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7981 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     8041 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     8101 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     8161 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     8221 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaatt
//
