LOCUS       sequence-139925-        7463 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 3 protein, C-terminally fused to the Fc 
            region of human IgG1 + 6X histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7463)
  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 7463)
  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-Fc-His
FEATURES             Location/Qualifiers
     source          1..7463
                     /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             4043..4060
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4086..4207
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4123..4142)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4177..4196
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4245..4262)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4416..5004)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4496..4515)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5175..5966,5967..6035))
                     /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     5798..5817
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6036..6140)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6208..6226
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6264..6286)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6288..6308)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6293..6623
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6474..6609
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6536..6555
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6760..6779
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6846..7301)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6983..7004)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7195..7214
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7427..7449
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7441..7458
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7442..7458
                     /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 atgatgaggc gagggagcct cgctcttgta ttcctggcct
     1741 tgtgaacgct gctctgggcc gagaggtgct ggcgtccagc acgtgcggga ggtcggccaa
     1801 tcgcgtctgc gattcctcgg acccgcagcg ggcacactct 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 ccaactctag acgcggcctg cggttgcccg cgcctcctgc ccggccggcg
     3181 ttacttgctg ctgggaggtg ggccgggggc tgcagctggg agcacagcgg gccggggaca
     3241 ggggctcagt gctgcccgtg gaagcctcgt gctgccttgg agagacgcct ggacccggcg
     3301 cctgcggagg ctgcagagga gagagcggcg ggggcgctgc gggaccggcg cgcctgacaa
     3361 aactcacaca tgcccaccgt gcccagcacc tgaactcctg gggggaccgt cagtcttcct
     3421 cttcccccca aaacccaagg acaccctcat gatctcccgg acccctgagg tcacatgcgt
     3481 ggtggtggac gtgagccacg aagaccctga ggtcaagttc aactggtacg tggacggcgt
     3541 ggaggtgcat aatgccaaga caaagccgcg ggaggagcag tacaacagca cgtaccgtgt
     3601 ggtcagcgtc ctcaccgtcc tgcaccagga ctggctgaat ggcaaggagt acaagtgcaa
     3661 ggtctccaac aaagccctcc cagcccccat cgagaaaacc atctccaaag ccaaagggca
     3721 gccccgagaa ccacaggtgt ataccctgcc cccatcccgg gatgagctga ccaagaacca
     3781 ggtcagcctg acctgcctgg tcaaaggctt ctatcccagc gacatcgccg tggagtggga
     3841 gagcaatggg cagccggaga acaactacaa gaccacgcct cccgtgctgg actccgacgg
     3901 ctccttcttc ctctacagca agctcaccgt ggacaagagc aggtggcagc aggggaacgt
     3961 cttctcatgc tccgtgatgc atgaggctct gcacaaccac tacacgcaga agagcctctc
     4021 cctgtctccg ggtaaaggcg gccatcatca ccatcaccat taatctagag atcttattaa
     4081 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     4141 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     4201 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     4261 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     4321 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     4381 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     4441 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     4501 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     4561 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     4621 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     4681 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     4741 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     4801 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     4861 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     4921 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     4981 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     5041 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     5101 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     5161 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     5221 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     5281 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     5341 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     5401 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     5461 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     5521 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     5581 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     5641 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     5701 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     5761 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     5821 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     5881 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     5941 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     6001 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     6061 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     6121 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     6181 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     6241 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     6301 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     6361 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     6421 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     6481 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     6541 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     6601 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     6661 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     6721 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     6781 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     6841 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     6901 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     6961 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     7021 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     7081 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     7141 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     7201 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     7261 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     7321 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     7381 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     7441 tgtaaaacga cggccagtga att
//
