LOCUS       sequence-139918-        7271 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 5, isoform z 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 7271)
  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 7271)
  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: Ntn5.z-Fc-His
FEATURES             Location/Qualifiers
     source          1..7271
                     /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             3851..3868
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3894..4015
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3931..3950)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3985..4004
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4053..4070)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4224..4812)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4304..4323)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4983..5774,5775..5843))
                     /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     5606..5625
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5844..5948)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6016..6034
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6072..6094)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6096..6116)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6101..6431
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6282..6417
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6344..6363
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6568..6587
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6654..7109)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6791..6812)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7003..7022
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7235..7257
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7249..7266
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7250..7266
                     /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 tgaccgacta
     1621 caggacactc ttttcttctc caggtgccgg ctctactgtg accacgccca ttaccctcag
     1681 cctcttgctc ctccttagcc aggccacctc agacccatgc tatgatccag gagggcgccc
     1741 tcgcttttgc ctcccaccag tgacccagtt ggttggcaag gcagcagccc cctgctctca
     1801 gacctgtgct cttcctgcag ccagccctgg ccctgcctgc aacagtagtc tgaccctgga
     1861 cctggacggc tccttcctct tgacatctgt gaccctgcgt ttctgcacag cgggacctcc
     1921 agccctggtt ctttctgctg cctgggccac tggggggccc tggaggccac tatggcgtag
     1981 gcctgcctgg cctggggctc tggggggccc caagaaggta acctttcact cccccccagg
     2041 ccccaagacc aggatagtgg ccagctacct ccgtgtggag ttcgggggaa aagcagggct
     2101 ggtgaccact ggagtaagag gccgctgcca gtgccacggc cacgctgccc gctgtgccac
     2161 tcgggcccag cccccacgct gccgctgccg ccaccacacc actggcccag ggtgtgagag
     2221 ctgtcgccca tcccatcgag actggccctg gcgacctgcc acaccccagc accctcaccc
     2281 gtgcttgccc tgttcctgca atcagcatgc ccgccgctgc cgattcaact ccgagctatt
     2341 caggttgtca ggtggccgta gtgggggtgt gtgtgagcgg tgccgccacc acacagctgg
     2401 gcggcactgt caccactgcc agccagggtt ctggagggac ccgagccagc ccatcactag
     2461 tcacaaggca tgcagggcct gccagtgcca cccaattgga gcaacgggag ggatgtgcaa
     2521 ccagaccagt ggccagtgct cctgcaagtt aggggtcaca ggcctgacat gcaatcgctg
     2581 tggtcctggc taccagcaga gccgctcacc caggatgccc tgccaacgaa ttccagaggc
     2641 aacaactacc cctgctacta cccctgttgc atctcgatca gacccacagt gtcaaggcta
     2701 ttgcaatgtc tccgtcagca gtgtgcacat gagccttcag agatactgtc agcaggatta
     2761 tgttctgcac gcccaggttt cagcgtcgtc gtcgcagccg tctgaggcgg tgggccccga
     2821 gtggtggcgg ctggccgtgc acgtgctggc tgtgttcaaa cagcgtgcgt ggcccgtgcg
     2881 ccgcggtggc caggaggcct gggtgccacg tgctgaccta atctgcggct gcctgcgcct
     2941 gcgccctggc gctgactacc tcctgctggg ccgtgctgcg cagacccacg acgacgacaa
     3001 ctatgaccct gcgcgcctca tcctgaaccg ccacggcctc gcgctgccct ggagaccacg
     3061 atgggcccgg ccgctgcggc gactgcagca gaaggagcgc ggaggtgcct gccgaggcct
     3121 gctgccaccc acacggagcc ctgggcccag gaacggcgcg cctgacaaaa ctcacacatg
     3181 cccaccgtgc ccagcacctg aactcctggg gggaccgtca gtcttcctct tccccccaaa
     3241 acccaaggac accctcatga tctcccggac ccctgaggtc acatgcgtgg tggtggacgt
     3301 gagccacgaa gaccctgagg tcaagttcaa ctggtacgtg gacggcgtgg aggtgcataa
     3361 tgccaagaca aagccgcggg aggagcagta caacagcacg taccgtgtgg tcagcgtcct
     3421 caccgtcctg caccaggact ggctgaatgg caaggagtac aagtgcaagg tctccaacaa
     3481 agccctccca gcccccatcg agaaaaccat ctccaaagcc aaagggcagc cccgagaacc
     3541 acaggtgtat accctgcccc catcccggga tgagctgacc aagaaccagg tcagcctgac
     3601 ctgcctggtc aaaggcttct atcccagcga catcgccgtg gagtgggaga gcaatgggca
     3661 gccggagaac aactacaaga ccacgcctcc cgtgctggac tccgacggct ccttcttcct
     3721 ctacagcaag ctcaccgtgg acaagagcag gtggcagcag gggaacgtct tctcatgctc
     3781 cgtgatgcat gaggctctgc acaaccacta cacgcagaag agcctctccc tgtctccggg
     3841 taaaggcggc catcatcacc atcaccatta atctagagat cttattaaag cagaacttgt
     3901 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3961 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4021 tctggtcgac tctagctaga ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4081 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4141 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4201 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4261 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4321 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4381 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     4441 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     4501 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     4561 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     4621 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     4681 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     4741 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     4801 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     4861 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     4921 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     4981 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5041 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5101 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5161 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5221 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5281 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5341 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     5401 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     5461 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     5521 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     5581 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     5641 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     5701 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     5761 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     5821 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     5881 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     5941 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6001 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6061 gacggtgaaa acctctgaca catgcagctc ccggaggaat gtgtgtcagt tagggtgtgg
     6121 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6181 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6241 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6301 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6361 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     6421 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cgtcccggag acggtcacag
     6481 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     6541 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     6601 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     6661 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     6721 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     6781 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     6841 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     6901 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     6961 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7021 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7081 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     7141 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7201 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7261 gccagtgaat t
//
