LOCUS       sequence-131167-        7871 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 5, isoform a 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 7871)
  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 7871)
  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-AP-His
FEATURES             Location/Qualifiers
     source          1..7871
                     /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             4451..4468
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4494..4615
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4531..4550)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4585..4604
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4653..4670)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4824..5412)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4904..4923)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5583..6374,6375..6443))
                     /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     6206..6225
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6444..6548)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6616..6634
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6672..6694)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6696..6716)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6701..7031
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6882..7017
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6944..6963
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7168..7187
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7254..7709)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7391..7412)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7603..7622
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7835..7857
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7849..7866
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7850..7866
                     /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 tggatcttcc ggaatcatcc cagttgagga
     3001 ggagaacccg gacttctgga accgcgaggc agccgaggcc ctgggtgccg ccaagaagct
     3061 gcagcctgca cagacagccg ccaagaacct catcatcttc ctgggcgatg ggatgggggt
     3121 gtctacggtg acagctgcca ggatcctaaa agggcagaag aaggacaaac tggggcctga
     3181 gatacccctg gccatggacc gcttcccata tgtggctctg tccaagacat acaatgtaga
     3241 caaacatgtg ccagacagtg gagccacagc cacggcctac ctgtgcgggg tcaagggcaa
     3301 cttccagacc attggcttga gtgcagccgc ccgctttaac cagtgcaaca cgacacgcgg
     3361 caacgaggtc atctccgtga tgaatcgggc caagaaagca gggaagtcag tgggagtggt
     3421 aaccaccaca cgagtgcagc acgcctcgcc agccggcacc tacgcccaca cggtgaaccg
     3481 caactggtac tcggacgccg acgtgcctgc ctcggcccgc caggaggggt gccaggacat
     3541 cgctacgcag ctcatctcca acatggacat tgacgtgatc ctaggtgggg gccgaaagta
     3601 catgtttcgc atgggaaccc cagaccctga gtacccagat gactacagcc aaggtgggac
     3661 caggctggac gggaagaatc tggtgcagga atggctggcg aagcgccagg gtgcccggta
     3721 cgtgtggaac cgcactgagc tcatgcaggc ttccctggac ccgtctgtgg cccatctcat
     3781 gggtctcttt gagcctggag acatgaaata cgagatccac cgagactcca cactggaccc
     3841 ctccctgatg gagatgacag aggctgccct gcgcctgctg agcaggaacc cccgcggctt
     3901 cttcctcttc gtggagggtg gtcgcatcga ccatggtcat catgaaagca gggcttaccg
     3961 ggcactgact gagacgatca tgttcgacga cgccattgag agggcgggcc agctcaccag
     4021 cgaggaggac acgctgagcc tcgtcactgc cgaccactcc cacgtcttct ccttcggagg
     4081 ctaccccctg cgagggagct ccatcttcgg gctggcccct ggcaaggccc gggacaggaa
     4141 ggcctacacg gtcctcctat acggaaacgg tccaggctat gtgctcaagg acggcgcccg
     4201 gccggatgtt accgagagcg agagcgggag ccccgagtat cggcagcagt cagcagtgcc
     4261 cctggacgaa gagacccacg caggcgagga cgtggcggtg ttcgcgcgcg gcccgcaggc
     4321 gcacctggtt cacggcgtgc aggagcagac cttcatagcg cacgtcatgg ccttcgccgc
     4381 ctgcctggag ccctacaccg cctgcgacct ggcgcccccc gccggcacca ccgacgccgc
     4441 gcacccgggt catcatcacc atcaccatta atctagagat cttattaaag cagaacttgt
     4501 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4561 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4621 tctggtcgac tctagctaga ctcttccgct tcctcgctca ctgactcgct gcgctcggtc
     4681 gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa
     4741 tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt
     4801 aaaaaggccg cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa
     4861 aatcgacgct caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt
     4921 ccccctggaa gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg
     4981 tccgcctttc tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc
     5041 agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc
     5101 gaccgctgcg ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta
     5161 tcgccactgg cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct
     5221 acagagttct tgaagtggtg gcctaactac ggctacacta gaagaacagt atttggtatc
     5281 tgcgctctgc tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa
     5341 caaaccaccg ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa
     5401 aaaggatctc aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa
     5461 aactcacgtt aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt
     5521 ttaaattaaa aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac
     5581 agttaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc
     5641 atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt accatctggc
     5701 cccagtgctg caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata
     5761 aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc
     5821 cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc
     5881 aacgttgttg ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca
     5941 ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa
     6001 gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca
     6061 ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt
     6121 tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt
     6181 tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg
     6241 ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga
     6301 tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc
     6361 agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg
     6421 acacggaaat gttgaatact catactcttc ctttttcaat attattgaag catttatcag
     6481 ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg
     6541 gttccgcgca catttccccg aaaagtgcca cctgacgtct aagaaaccat tattatcatg
     6601 acattaacct ataaaaatag gcgtatcacg aggccctttc gtctcgcgcg tttcggtgat
     6661 gacggtgaaa acctctgaca catgcagctc ccggaggaat gtgtgtcagt tagggtgtgg
     6721 aaagtcccca ggctccccag gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     6781 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     6841 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     6901 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     6961 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     7021 gcttttgcaa aaagctcccg ggagcttgta tatccatttt cgtcccggag acggtcacag
     7081 cttgtctgta agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg
     7141 gcgggtgtcg gggctggctt aactatgcgg catcagagca gattgtactg agagtgcacc
     7201 atatgcggtg tgaaataccg cacagatgcg taaggagaaa ataccgcatc aggaaattgt
     7261 aagcgttaat attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa
     7321 ccaataggcc gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt
     7381 gagtgttgtt ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa
     7441 agggcgaaaa accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag
     7501 ttttttgggg tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt
     7561 tagagcttga cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg
     7621 agcgggcgct agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc
     7681 cgcgcttaat gcgccgctac agggcgcgtc cattcgccat tcaggctgcg caactgttgg
     7741 gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg gggatgtgct
     7801 gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg taaaacgacg
     7861 gccagtgaat t
//
