LOCUS       sequence-138919-        7610 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 4 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 7610)
  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 7610)
  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: Ntn4-Fc-His
FEATURES             Location/Qualifiers
     source          1..7610
                     /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             4190..4207
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4233..4354
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4270..4289)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4324..4343
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4392..4409)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4563..5151)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4643..4662)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5322..6113,6114..6182))
                     /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     5945..5964
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6183..6287)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6355..6373
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6411..6433)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6435..6455)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6440..6770
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6621..6756
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6683..6702
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6907..6926
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6993..7448)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7130..7151)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7342..7361
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7574..7596
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7588..7605
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7589..7605
                     /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 tggggagctg
     1621 cgcacggctg ctgctgctct ggggctgctc cgcggtggcc gcaggcttga atggagtagc
     1681 cggagcgaac tcccgctgtg agaaggcatg caaccctcgc atgggaaact tggctttggg
     1741 aagaaagctc cgggcagaca ctatgtgtgg ccagaacgcc accgaactct tctgcttcta
     1801 cagtgagaat gctgacctca cttgccggca gcccaagtgt gataaatgca acgctgccca
     1861 ttctcaccta gctcacccac cctctgccat ggcagactca tccttcaggt ttccccggac
     1921 atggtggcag tctgcagagg atgtgcacag ggaaaagatt cagctagacc tggaagcaga
     1981 attctacttc actcacctaa ttatggtgtt caagtctccc aggcctgcag ccatggtgct
     2041 ggaccggtcc caggactttg ggaagacctg gaagccttac aagtactttg caacaaactg
     2101 ctcggctact tttggcctgg aagatgatgt tgtcaagaag ggagctattt gcacgtctag
     2161 atactcaaat cctttcccgt gcaccggagg agaggttatt ttcagagccc tgtcaccacc
     2221 atacgacata gaaaaccctt acagtgccaa agtgcaggag cagctgaaga tcaccaacct
     2281 ccgagtgcgg ctgctcaagc gacagtcctg cccttgtcag ataaacgacc tgaacgcaaa
     2341 acctcaccat tttatgcact acgcagtcta tgacttcatc gtcaagggca gctgcttctg
     2401 caacggccac gctgaccagt gcttacctgt ggagggcttc agacccatca aggccccggg
     2461 agcgttccac gtggtccacg ggaggtgtat gtgtaagcac aacacagcag gcagccactg
     2521 ccagcactgt gcaccattgt acaatgaccg gccctgggag gcagcggacg gcagaacagg
     2581 ggctcctaac gaatgcagaa cttgcaagtg caatgggcac gcggacacct gtcacttcga
     2641 cgtcaacgtg tgggaggcgt cggggaaccg cagcggcggt gtctgcaaca actgtcagca
     2701 caacactgag ggtcagcact gtcaacgctg taagcccggt ttctaccgcg acctcagaag
     2761 acccttctct gcccctgacg cttgcaaagc gtgttcctgc cacccggttg gatcggcgat
     2821 ccttcctttc agctcagtga ccttctgcga ccccagcaat ggtgactgcc cctgcaagcc
     2881 tggggtggcg gggccacatt gcgacagatg catggtggga tactggggct ttggagacta
     2941 cggctgcaga ccttgcgatt gtgcggggag ctgcgacccg ctcacgggag actgcatcag
     3001 cagtaacgct gatgtagact ggtaccacga agtccccgcc tttcactcga tgcacaataa
     3061 gagtgagccc agctgggaat gggaggatga gcaaggattt tctgccctcc gacactcagg
     3121 taaatgtgaa tgtaaggaac aggtgttagg aaaccccaaa gccttctgtg gaatgaagta
     3181 ttcatatgtg ttaaaaatca agatcttatc agcccacgac aaaggctccc atgccgaagt
     3241 caatgtgaag attaagaaag tcttaaagtc caccaaactg aagatcttac gaggcaagag
     3301 aacgctatac ccagagtcct ggactaacag aggctgcacc tgtccaatcc tcaatccagg
     3361 attggagtac ctggtcgctg gccacgagga cgtaagaaca ggcaaattaa ttgtgaatat
     3421 gaaaagcttt gtccagcact ggaaaccagc tcttggcaga agagtcatgc acatcttaaa
     3481 aagagactgc gtgggcgcgc ctgacaaaac tcacacatgc ccaccgtgcc cagcacctga
     3541 actcctgggg ggaccgtcag tcttcctctt ccccccaaaa cccaaggaca ccctcatgat
     3601 ctcccggacc cctgaggtca catgcgtggt ggtggacgtg agccacgaag accctgaggt
     3661 caagttcaac tggtacgtgg acggcgtgga ggtgcataat gccaagacaa agccgcggga
     3721 ggagcagtac aacagcacgt accgtgtggt cagcgtcctc accgtcctgc accaggactg
     3781 gctgaatggc aaggagtaca agtgcaaggt ctccaacaaa gccctcccag cccccatcga
     3841 gaaaaccatc tccaaagcca aagggcagcc ccgagaacca caggtgtata ccctgccccc
     3901 atcccgggat gagctgacca agaaccaggt cagcctgacc tgcctggtca aaggcttcta
     3961 tcccagcgac atcgccgtgg agtgggagag caatgggcag ccggagaaca actacaagac
     4021 cacgcctccc gtgctggact ccgacggctc cttcttcctc tacagcaagc tcaccgtgga
     4081 caagagcagg tggcagcagg ggaacgtctt ctcatgctcc gtgatgcatg aggctctgca
     4141 caaccactac acgcagaaga gcctctccct gtctccgggt aaaggcggcc atcatcacca
     4201 tcaccattaa tctagagatc ttattaaagc agaacttgtt tattgcagct tataatggtt
     4261 acaaataaag caatagcatc acaaatttca caaataaagc atttttttca ctgcattcta
     4321 gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggtcgact ctagctagac
     4381 tcttccgctt cctcgctcac tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta
     4441 tcagctcact caaaggcggt aatacggtta tccacagaat caggggataa cgcaggaaag
     4501 aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg
     4561 tttttccata ggctccgccc ccctgacgag catcacaaaa atcgacgctc aagtcagagg
     4621 tggcgaaacc cgacaggact ataaagatac caggcgtttc cccctggaag ctccctcgtg
     4681 cgctctcctg ttccgaccct gccgcttacc ggatacctgt ccgcctttct cccttcggga
     4741 agcgtggcgc tttctcatag ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc
     4801 tccaagctgg gctgtgtgca cgaacccccc gttcagcccg accgctgcgc cttatccggt
     4861 aactatcgtc ttgagtccaa cccggtaaga cacgacttat cgccactggc agcagccact
     4921 ggtaacagga ttagcagagc gaggtatgta ggcggtgcta cagagttctt gaagtggtgg
     4981 cctaactacg gctacactag aagaacagta tttggtatct gcgctctgct gaagccagtt
     5041 accttcggaa aaagagttgg tagctcttga tccggcaaac aaaccaccgc tggtagcggt
     5101 ggtttttttg tttgcaagca gcagattacg cgcagaaaaa aaggatctca agaagatcct
     5161 ttgatctttt ctacggggtc tgacgctcag tggaacgaaa actcacgtta agggattttg
     5221 gtcatgagat tatcaaaaag gatcttcacc tagatccttt taaattaaaa atgaagtttt
     5281 aaatcaatct aaagtatata tgagtaaact tggtctgaca gttaccaatg cttaatcagt
     5341 gaggcaccta tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc
     5401 gtgtagataa ctacgatacg ggagggctta ccatctggcc ccagtgctgc aatgataccg
     5461 cgagacccac gctcaccggc tccagattta tcagcaataa accagccagc cggaagggcc
     5521 gagcgcagaa gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg
     5581 gaagctagag taagtagttc gccagttaat agtttgcgca acgttgttgc cattgctaca
     5641 ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga
     5701 tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct
     5761 ccgatcgttg tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg
     5821 cataattctc ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca
     5881 accaagtcat tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata
     5941 cgggataata ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct
     6001 tcggggcgaa aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact
     6061 cgtgcaccca actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa
     6121 acaggaaggc aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc
     6181 atactcttcc tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga
     6241 tacatatttg aatgtattta gaaaaataaa caaatagggg ttccgcgcac atttccccga
     6301 aaagtgccac ctgacgtcta agaaaccatt attatcatga cattaaccta taaaaatagg
     6361 cgtatcacga ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     6421 atgcagctcc cggaggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagg
     6481 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccaggtgt ggaaagtccc
     6541 caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca gcaaccatag
     6601 tcccgcccct aactccgccc atcccgcccc taactccgcc cagttccgcc cattctccgc
     6661 cccatggctg actaattttt tttatttatg cagaggccga ggccgcctct gcctctgagc
     6721 tattccagaa gtagtgagga ggcttttttg gaggcctagg cttttgcaaa aagctcccgg
     6781 gagcttgtat atccattttc gtcccggaga cggtcacagc ttgtctgtaa gcggatgccg
     6841 ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta
     6901 actatgcggc atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc
     6961 acagatgcgt aaggagaaaa taccgcatca ggaaattgta agcgttaata ttttgttaaa
     7021 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     7081 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     7141 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     7201 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     7261 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     7321 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     7381 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     7441 gggcgcgtcc attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc
     7501 tcttcgctat tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta
     7561 acgccagggt tttcccagtc acgacgttgt aaaacgacgg ccagtgaatt
//
