LOCUS       sequence-139088-        7079 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 5, isoform a 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 7079)
  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 7079)
  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.a-Fc-His
FEATURES             Location/Qualifiers
     source          1..7079
                     /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             3659..3676
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    3702..3823
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3739..3758)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3793..3812
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3861..3878)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4032..4620)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4112..4131)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4791..5582,5583..5651))
                     /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     5414..5433
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5652..5756)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5824..5842
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(5880..5902)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5904..5924)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5909..6239
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6090..6225
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6152..6171
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6376..6395
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6462..6917)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(6599..6620)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     6811..6830
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7043..7065
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7057..7074
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7058..7074
                     /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 tgccagtgcc acccaattgg agcaacggga gggatgtgca accagaccag
     2341 tggccagtgc tcctgcaagt taggggtcac aggcctgaca tgcaatcgct gtggtcctgg
     2401 ctaccagcag agccgctcac ccaggatgcc ctgccaacga attccagagg caacaactac
     2461 ccctgctact acccctgttg catctcgatc agacccacag tgtcaaggct attgcaatgt
     2521 ctccgtcagc agtgtgcaca tgagccttca gagatactgt cagcaggatt atgttctgca
     2581 cgcccaggtt tcagcgtcgt cgtcgcagcc gtctgaggcg gtgggccccg agtggtggcg
     2641 gctggccgtg cacgtgctgg ctgtgttcaa acagcgtgcg tggcccgtgc gccgcggtgg
     2701 ccaggaggcc tgggtgccac gtgctgacct aatctgcggc tgcctgcgcc tgcgccctgg
     2761 cgctgactac ctcctgctgg gccgtgctgc gcagacccac gacgacaact atgaccctgc
     2821 gcgcctcatc ctgaaccgcc acggcctcgc gctgccctgg agaccacgat gggcccggcc
     2881 gctgcggcga ctgcagcaga aggagcgcgg aggtgcctgc cgaggcctgc tgccacccac
     2941 acggagccct gggcccagga acggcgcgcc tgacaaaact cacacatgcc caccgtgccc
     3001 agcacctgaa ctcctggggg gaccgtcagt cttcctcttc cccccaaaac ccaaggacac
     3061 cctcatgatc tcccggaccc ctgaggtcac atgcgtggtg gtggacgtga gccacgaaga
     3121 ccctgaggtc aagttcaact ggtacgtgga cggcgtggag gtgcataatg ccaagacaaa
     3181 gccgcgggag gagcagtaca acagcacgta ccgtgtggtc agcgtcctca ccgtcctgca
     3241 ccaggactgg ctgaatggca aggagtacaa gtgcaaggtc tccaacaaag ccctcccagc
     3301 ccccatcgag aaaaccatct ccaaagccaa agggcagccc cgagaaccac aggtgtatac
     3361 cctgccccca tcccgggatg agctgaccaa gaaccaggtc agcctgacct gcctggtcaa
     3421 aggcttctat cccagcgaca tcgccgtgga gtgggagagc aatgggcagc cggagaacaa
     3481 ctacaagacc acgcctcccg tgctggactc cgacggctcc ttcttcctct acagcaagct
     3541 caccgtggac aagagcaggt ggcagcaggg gaacgtcttc tcatgctccg tgatgcatga
     3601 ggctctgcac aaccactaca cgcagaagag cctctccctg tctccgggta aaggcggcca
     3661 tcatcaccat caccattaat ctagagatct tattaaagca gaacttgttt attgcagctt
     3721 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     3781 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggtcgactc
     3841 tagctagact cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     3901 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     3961 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     4021 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     4081 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     4141 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     4201 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     4261 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     4321 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     4381 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     4441 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     4501 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     4561 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     4621 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     4681 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     4741 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     4801 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     4861 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     4921 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     4981 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     5041 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     5101 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     5161 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     5221 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     5281 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     5341 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     5401 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     5461 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     5521 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     5581 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     5641 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     5701 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     5761 tttccccgaa aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat
     5821 aaaaataggc gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac
     5881 ctctgacaca tgcagctccc ggaggaatgt gtgtcagtta gggtgtggaa agtccccagg
     5941 ctccccaggc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg
     6001 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6061 caaccatagt cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc
     6121 attctccgcc ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg
     6181 cctctgagct attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa
     6241 agctcccggg agcttgtata tccattttcg tcccggagac ggtcacagct tgtctgtaag
     6301 cggatgccgg gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg
     6361 gctggcttaa ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg
     6421 aaataccgca cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat
     6481 tttgttaaaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
     6541 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc
     6601 agtttggaac aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac
     6661 cgtctatcag ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc
     6721 gaggtgccgt aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg
     6781 gggaaagccg gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag
     6841 ggcgctggca agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc
     6901 gccgctacag ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg
     6961 gtgcgggcct cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta
     7021 agttgggtaa cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
