LOCUS       sequence-131244-        7457 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Unc5D, isoform y 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 7457)
  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 7457)
  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: Unc5d.y-AP-His
FEATURES             Location/Qualifiers
     source          1..7457
                     /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             4037..4054
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4080..4201
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4117..4136)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4171..4190
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4239..4256)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4410..4998)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4490..4509)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5169..5960,5961..6029))
                     /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     5792..5811
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6030..6134)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6202..6220
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6258..6280)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6282..6302)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6287..6617
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6468..6603
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6530..6549
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6754..6773
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6840..7295)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6977..6998)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7189..7208
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7421..7443
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7435..7452
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7436..7452
                     /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 tggggacagg
     1621 ggctgcagac ggcagccgcg gggcgcgccg gtggctccca tggctaggac tctttttctg
     1681 ggcggcgggg gctgcggctg cccgaggagc cgatggcagt gaaattcttc ccgactctat
     1741 cccatctgcc cctggaacgc tgcctcattt catcgaggag cccgaagatg catacatcat
     1801 caagagcaac ccgattgcac tcaggtgcaa agcaaggcca gccatgcaga tcttcttcaa
     1861 atgcaatggg gagtgggtcc atcagaatga acatgtgtcc gaggagagcc tggatgagag
     1921 ctcaggcttg aaggtccggg aagtgtttat caatgtcacc aggcagcaag tggaggactt
     1981 ccatgggcca gaggactact ggtgccagtg tgtagcgtgg agccacctgg gaacgtccaa
     2041 gagtaggaag gcatctgtgc gcatagccta tttacggaaa aactttgaac aagatccaca
     2101 aggcagggag gttcctattg aaggcatgat cgtgctgcat tgtcgcccgc cagagggagt
     2161 tccagcagca gaggtggaat ggctgaaaaa tgaggagccc atcgactccg aacaggatga
     2221 gaacattgac accagggctg accataactt gatcatcaga caggcccggc tgtcagactc
     2281 agggaattac acctgtatgg cagccaacat cgtggccaag aggaggagcc tctcagcaac
     2341 tgtggtggtc tacgtggatg gcagctggga agtgtggagt gaatggtcag tctgcagccc
     2401 agagtgtgaa catcttcgta tccgtgagtg cacagctcca cccccaagaa atgggggcaa
     2461 gttctgtgaa ggtctcagcc aggaatctga aaactgcaca gatggtctct gcattctagg
     2521 atggagtagg agaggcattg agaatgccag cactagtgga tcttccggaa tcatcccagt
     2581 tgaggaggag aacccggact tctggaaccg cgaggcagcc gaggccctgg gtgccgccaa
     2641 gaagctgcag cctgcacaga cagccgccaa gaacctcatc atcttcctgg gcgatgggat
     2701 gggggtgtct acggtgacag ctgccaggat cctaaaaggg cagaagaagg acaaactggg
     2761 gcctgagata cccctggcca tggaccgctt cccatatgtg gctctgtcca agacatacaa
     2821 tgtagacaaa catgtgccag acagtggagc cacagccacg gcctacctgt gcggggtcaa
     2881 gggcaacttc cagaccattg gcttgagtgc agccgcccgc tttaaccagt gcaacacgac
     2941 acgcggcaac gaggtcatct ccgtgatgaa tcgggccaag aaagcaggga agtcagtggg
     3001 agtggtaacc accacacgag tgcagcacgc ctcgccagcc ggcacctacg cccacacggt
     3061 gaaccgcaac tggtactcgg acgccgacgt gcctgcctcg gcccgccagg aggggtgcca
     3121 ggacatcgct acgcagctca tctccaacat ggacattgac gtgatcctag gtggaggccg
     3181 aaagtacatg tttcgcatgg gaaccccaga ccctgagtac ccagatgact acagccaagg
     3241 tgggaccagg ctggacggga agaatctggt gcaggaatgg ctggcgaagc gccagggtgc
     3301 ccggtatgtg tggaaccgca ctgagctcat gcaggcttcc ctggacccgt ctgtgaccca
     3361 tctcatgggt ctctttgagc ctggagacat gaaatacgag atccaccgag actccacact
     3421 ggacccctcc ctgatggaga tgacagaggc tgccctgcgc ctgctgagca ggaacccccg
     3481 cggcttcttc ctcttcgtgg agggtggtcg catcgaccat ggtcatcatg aaagcagggc
     3541 ttaccgggca ctgactgaga cgatcatgtt cgacgacgcc attgagaggg cgggccagct
     3601 caccagcgag gaggacacgc tgagcctcgt cactgccgac cactcccacg tcttctcctt
     3661 cggaggctac cccctgcgag ggagctccat cttcgggctg gcccctggca aggcccggga
     3721 caggaaggcc tacacggtcc tcctatacgg aaacggtcca ggctatgtgc tcaaggacgg
     3781 cgcccggccg gatgttaccg agagcgagag cgggagcccc gagtatcggc agcagtcagc
     3841 agtgcccctg gacgaagaga cccacgcagg cgaggacgtg gcggtgttcg cgcgcggccc
     3901 gcaggcgcac ctggttcacg gcgtgcagga gcagaccttc atagcgcacg tcatggcctt
     3961 cgccgcctgc ctggagccct acaccgcctg cgacctggcg ccccccgccg gcaccaccga
     4021 cgccgcgcac ccgggtcatc atcaccatca ccattaatct agagatctta ttaaagcaga
     4081 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4141 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4201 atcatgtctg gtcgactcta gctagactct tccgcttcct cgctcactga ctcgctgcgc
     4261 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4321 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4381 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4441 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4501 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4561 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     4621 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     4681 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     4741 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     4801 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     4861 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     4921 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     4981 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5041 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5101 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5161 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5221 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5281 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5341 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5401 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5461 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5521 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     5581 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     5641 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     5701 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     5761 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     5821 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     5881 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     5941 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6001 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6061 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6121 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6181 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6241 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg aggaatgtgt gtcagttagg
     6301 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     6361 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6421 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6481 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6541 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     6601 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgtc ccggagacgg
     6661 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     6721 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     6781 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     6841 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     6901 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     6961 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7021 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7081 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7141 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7201 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7261 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     7321 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7381 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7441 acgacggcca gtgaatt
//
