LOCUS       sequence-135596-        8255 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 3 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 8255)
  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 8255)
  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: Ntn3-AP-His
FEATURES             Location/Qualifiers
     source          1..8255
                     /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)"
     CDS             4835..4852
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4878..4999
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4915..4934)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4969..4988
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5037..5054)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5208..5796)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5288..5307)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5967..6758,6759..6827))
                     /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     6590..6609
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6828..6932)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7000..7018
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7056..7078)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7080..7100)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7085..7415
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7266..7401
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7328..7347
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7552..7571
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7638..8093)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7775..7796)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7987..8006
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8219..8241
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8233..8250
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8234..8250
                     /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 tgcccacctg
     1621 gctctggggg ctgctgctga ccgcgggcac gctctccgct gcactgagcc cagggctgcc
     1681 ggcctctgcc gacccctgct atgatgaagc gagggagcct cgctcttgta ttcctggcct
     1741 tgtgaacgct gctctgggcc gagaggtgct ggcgtccagc acgtgcggga ggtcggccaa
     1801 tcgcgtctgc gattcctcgg acccgcagcg ggctcactct gcagacctcc tgacctctgc
     1861 tccgggcact gcaagtcctc tctgttggcg ctccgatttg ctgcaacagg cacctttcaa
     1921 cgtaaccctc acagtgcccc tggggaaggc ttttgagctg gtcttcgtga gcctgcgctt
     1981 ctgctcagct cctccaacct ccgtggccct gcttaagtcg caggaccatg gccgcagctg
     2041 ggtccccttg ggcttcttct cttccagctg taccctggac tatggccgtc tgcctgctcc
     2101 tgctgatggc ccttctggtc cagggccaga agccctctgc tttccagccc cccaggctca
     2161 gcctgatggt ggaggccttc tggccttcag tgtgcaggat ggcagcccac agggcctgga
     2221 tctggacaac agccccgtgc tccaagactg ggtgactgcc acagatattc gcatagtact
     2281 cacaaggcct gccattcagg gagacaccag ggacggtggg gtgacagtcc cctactccta
     2341 ctcagccact gagcttcagg tgggaggtcg atgcaagtgc aatgggcatg cctcacggtg
     2401 tctgttggac acccatggcc acctggtctg cgactgccag catggtacag agggccctga
     2461 ttgcagccgc tgcaagccct tctactgcga caggccatgg cagcgggcta cagggcagga
     2521 agcccacgct tgccttgctt gctcctgcaa cggccatgcg cgaagatgcc gcttcaacat
     2581 ggagctctac cgactgtccg gccgccgcag tgggggcgtg tgcctcaact gccggcacaa
     2641 tacagctggt cgtcactgcc actactgccg ggagggcttc tatcgtgatc caggccgtgt
     2701 cctgagtgac cgtcgtgctt gcagagcttg tgactgccac ccagttggtg ctgctggcaa
     2761 aacctgtaac cagaccacag gccagtgtcc ctgtaaggat ggtgttactg gcctcacctg
     2821 taaccgctgt gccccaggtt tccagcagag ccgttctcct gtggcacctt gcgttaagac
     2881 tcctgtccct ggacccaccg aagaaagcag tcctgtggag ccacaggact gtgagtcaca
     2941 ttgcagacct gcgcgtggca gttaccgaat cagcctgaag aagttctgcc ggaaggacta
     3001 tgcggtgcag gtggcagtgg gtgcacgcgg tgaggcccgc ggctcgtgga cacgctttcc
     3061 ggtagcggtg cttgctgtgt tccgcagcgg cgaggaacgc gctcgacgcg ggagcagcgc
     3121 gctgtgggta ccaaccctag acgcggcctg cggttgcccg cgcctcctgc ctggccggcg
     3181 ttacttgctg ctgggaggtg ggccgggggc tgcagctggg agcacagcgg gccggggaca
     3241 ggggctcagt gctgcccgtg gaagcctcgt gctgccttgg agagacgcct ggacccggcg
     3301 cctgcggagg ctgcagagga gagagcggcg ggggcgctgc gggaccggcg cgcctggatc
     3361 ttccggaatc atcccagttg aggaggagaa cccggacttc tggaaccgcg aggcagccga
     3421 ggccctgggt gccgccaaga agctgcagcc tgcacagaca gccgccaaga acctcatcat
     3481 cttcctgggc gatgggatgg gggtgtctac ggtgacagct gccaggatcc taaaagggca
     3541 gaagaaggac aaactggggc ctgagatacc cctggccatg gaccgcttcc catatgtggc
     3601 tctgtccaag acatacaatg tagacaaaca tgtgccagac agtggagcca cagccacggc
     3661 ctacctgtgc ggggtcaagg gcaacttcca gaccattggc ttgagtgcag ccgcccgctt
     3721 taaccagtgc aacacgacac gcggcaacga ggtcatctcc gtgatgaatc gggccaagaa
     3781 agcagggaag tcagtgggag tggtaaccac cacacgagtg cagcacgcct cgccagccgg
     3841 cacctacgcc cacacggtga accgcaactg gtactcggac gccgacgtgc ctgcctcggc
     3901 ccgccaggag gggtgccagg acatcgctac gcagctcatc tccaacatgg acattgacgt
     3961 gatcctaggt gggggccgaa agtacatgtt tcgcatggga accccagacc ctgagtaccc
     4021 agatgactac agccaaggtg ggaccaggct ggacgggaag aatctggtgc aggaatggct
     4081 ggcgaagcgc cagggtgccc ggtatgtgtg gaaccgcact gagctcatgc aggcttccct
     4141 ggacccgtct gtgacccatc tcatgggtct ctttgagcct ggagacatga aatacgagat
     4201 ccaccgagac tccacactgg acccctccct gatggagatg acagaggctg ccctgcgcct
     4261 gctgagcagg aacccccgcg gcttcttcct cttcgtggag ggtggtcgca tcgaccatgg
     4321 tcatcatgaa agcagggctt accgggcact gactgagacg atcatgttcg acgacgccat
     4381 tgagagggcg ggccagctca ccagcgagga ggacacgctg agcctcgtca ctgccgacca
     4441 ctcccacgtc ttctccttcg gaggctaccc cctgcgaggg agctccatct tcgggctggc
     4501 ccctggcaag gcccgggaca ggaaggccta cacggtcctc ctatacggaa acggtccagg
     4561 ctatgtgctc aaggacggcg cccggccgga tgttaccgag agcgagagcg ggagccccga
     4621 gtatcggcag cagtcagcag tgcccctgga cgaagagacc cacgcaggcg aggacgtggc
     4681 ggtgttcgcg cgcggcccgc aggcgcacct ggttcacggc gtgcaggagc agaccttcat
     4741 agcgcacgtc atggccttcg ccgcctgcct ggagccctac accgcctgcg acctggcgcc
     4801 ccccgccggc accaccgacg ccgcgcaccc gggtcatcat caccatcacc attaatctag
     4861 agatcttatt aaagcagaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     4921 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     4981 aactcatcaa tgtatcttat catgtctggt cgactctagc tagactcttc cgcttcctcg
     5041 ctcactgact cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag
     5101 gcggtaatac ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa
     5161 ggccagcaaa aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc
     5221 cgcccccctg acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca
     5281 ggactataaa gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg
     5341 accctgccgc ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct
     5401 catagctcac gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt
     5461 gtgcacgaac cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag
     5521 tccaacccgg taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc
     5581 agagcgaggt atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac
     5641 actagaagaa cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga
     5701 gttggtagct cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc
     5761 aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg
     5821 gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca
     5881 aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt
     5941 atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca
     6001 gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg
     6061 atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca
     6121 ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt
     6181 cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt
     6241 agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca
     6301 cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca
     6361 tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga
     6421 agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact
     6481 gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga
     6541 gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg
     6601 ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc
     6661 tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga
     6721 tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat
     6781 gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt
     6841 caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt
     6901 atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac
     6961 gtctaagaaa ccattattat catgacatta acctataaaa ataggcgtat cacgaggccc
     7021 tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca gctcccggag
     7081 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccaggcaggc agaagtatgc
     7141 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     7201 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     7261 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     7321 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     7381 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     7441 ttttcgtccc ggagacggtc acagcttgtc tgtaagcgga tgccgggagc agacaagccc
     7501 gtcagggcgc gtcagcgggt gttggcgggt gtcggggctg gcttaactat gcggcatcag
     7561 agcagattgt actgagagtg caccatatgc ggtgtgaaat accgcacaga tgcgtaagga
     7621 gaaaataccg catcaggaaa ttgtaagcgt taatattttg ttaaaattcg cgttaaattt
     7681 ttgttaaatc agctcatttt ttaaccaata ggccgaaatc ggcaaaatcc cttataaatc
     7741 aaaagaatag accgagatag ggttgagtgt tgttccagtt tggaacaaga gtccactatt
     7801 aaagaacgtg gactccaacg tcaaagggcg aaaaaccgtc tatcagggcg atggcccact
     7861 acgtgaacca tcaccctaat caagtttttt ggggtcgagg tgccgtaaag cactaaatcg
     7921 gaaccctaaa gggagccccc gatttagagc ttgacgggga aagccggcga acgtggcgag
     7981 aaaggaaggg aagaaagcga aaggagcggg cgctagggcg ctggcaagtg tagcggtcac
     8041 gctgcgcgta accaccacac ccgccgcgct taatgcgccg ctacagggcg cgtccattcg
     8101 ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc gggcctcttc gctattacgc
     8161 cagctggcga aagggggatg tgctgcaagg cgattaagtt gggtaacgcc agggttttcc
     8221 cagtcacgac gttgtaaaac gacggccagt gaatt
//
