LOCUS       sequence-135806-        8060 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 5, isoform z 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 8060)
  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 8060)
  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-AP-His
FEATURES             Location/Qualifiers
     source          1..8060
                     /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             4640..4657
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4683..4804
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4720..4739)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4774..4793
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4842..4859)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5013..5601)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5093..5112)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5772..6563,6564..6632))
                     /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     6395..6414
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6633..6737)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6805..6823
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6861..6883)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6885..6905)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6890..7220
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7071..7206
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7133..7152
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7357..7376
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7443..7898)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7580..7601)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7792..7811
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8024..8046
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8038..8055
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8039..8055
                     /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 cactactgcc 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 acgacaacta
     3001 tgaccctgcg cgcctcatcc tgaaccgcca cggcctcgcg ctgccctgga gaccacgatg
     3061 ggcccggccg ctgcggcgac tgcagcagaa ggagcgcgga ggtgcctgcc gaggcctgct
     3121 gccacccaca cggagccctg ggcccaggaa cggcgcgcct ggatcttccg gaatcatccc
     3181 agttgaggag gagaacccgg acttctggaa ccgcgaggca gccgaggccc tgggtgccgc
     3241 caagaagctg cagcctgcac agacagccgc caagaacctc atcatcttcc tgggcgatgg
     3301 gatgggggtg tctacggtga cagctgccag gatcctaaaa gggcagaaga aggacaaact
     3361 ggggcctgag atacccctgg ccatggaccg cttcccatat gtggctctgt ccaagacata
     3421 caatgtagac aaacatgtgc cagacagtgg agccacagcc acggcctacc tgtgcggggt
     3481 caagggcaac ttccagacca ttggcttgag tgcagccgcc cgctttaacc agtgcaacac
     3541 gacacgcggc aacgaggtca tctccgtgat gaatcgggcc aagaaagcag ggaagtcagt
     3601 gggagtggta accaccacac gagtgcagca cgcctcgcca gccggcacct acgcccacac
     3661 ggtgaaccgc aactggtact cggacgccga cgtgcctgcc tcggcccgcc aggaggggtg
     3721 ccaggacatc gctacgcagc tcatctccaa catggacatt gacgtgatcc taggtggggg
     3781 ccgaaagtac atgtttcgca tgggaacccc agaccctgag tacccagatg actacagcca
     3841 aggtgggacc aggctggacg ggaagaatct ggtgcaggaa tggctggcga agcgccaggg
     3901 tgcccggtat gtgtggaacc gcactgagct catgcaggct tccctggacc cgtctgtgac
     3961 ccatctcatg ggtctctttg agcctggaga catgaaatac gagatccacc gagactccac
     4021 actggacccc tccctgatgg agatgacaga ggctgccctg cgcctgctga gcaggaaccc
     4081 ccgcggcttc ttcctcttcg tggagggtgg tcgcatcgac catggtcatc atgaaagcag
     4141 ggcttaccgg gcactgactg agacgatcat gttcgacgac gccattgaga gggcgggcca
     4201 gctcaccagc gaggaggaca cgctgagcct cgtcactgcc gaccactccc acgtcttctc
     4261 cttcggaggc taccccctgc gagggagctc catcttcggg ctggcccctg gcaaggcccg
     4321 ggacaggaag gcctacacgg tcctcctata cggaaacggt ccaggctatg tgctcaagga
     4381 cggcgcccgg ccggatgtta ccgagagcga gagcgggagc cccgagtatc ggcagcagtc
     4441 agcagtgccc ctggacgaag agacccacgc aggcgaggac gtggcggtgt tcgcgcgcgg
     4501 cccgcaggcg cacctggttc acggcgtgca ggagcagacc ttcatagcgc acgtcatggc
     4561 cttcgccgcc tgcctggagc cctacaccgc ctgcgacctg gcgccccccg ccggcaccac
     4621 cgacgccgcg cacccgggtc atcatcacca tcaccattaa tctagagatc ttattaaagc
     4681 agaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc acaaatttca
     4741 caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc atcaatgtat
     4801 cttatcatgt ctggtcgact ctagctagac tcttccgctt cctcgctcac tgactcgctg
     4861 cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta
     4921 tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc
     4981 aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag
     5041 catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac
     5101 caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc
     5161 ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag ctcacgctgt
     5221 aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc
     5281 gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga
     5341 cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta
     5401 ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag aagaacagta
     5461 tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga
     5521 tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg
     5581 cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag
     5641 tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc
     5701 tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact
     5761 tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt
     5821 cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta
     5881 ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta
     5941 tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc
     6001 gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat
     6061 agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt
     6121 atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg
     6181 tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca
     6241 gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta
     6301 agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg
     6361 cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact
     6421 ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg
     6481 ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt
     6541 actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga
     6601 ataagggcga cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc
     6661 atttatcagg gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa
     6721 caaatagggg ttccgcgcac atttccccga aaagtgccac ctgacgtcta agaaaccatt
     6781 attatcatga cattaaccta taaaaatagg cgtatcacga ggccctttcg tctcgcgcgt
     6841 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggaggaatg tgtgtcagtt
     6901 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     6961 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     7021 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     7081 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     7141 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     7201 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc gtcccggaga
     7261 cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag
     7321 cgggtgttgg cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga
     7381 gagtgcacca tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca
     7441 ggaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
     7501 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
     7561 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
     7621 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
     7681 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
     7741 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
     7801 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
     7861 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc attcgccatt caggctgcgc
     7921 aactgttggg aagggcgatc ggtgcgggcc tcttcgctat tacgccagct ggcgaaaggg
     7981 ggatgtgctg caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt
     8041 aaaacgacgg ccagtgaatt
//
