LOCUS       sequence-131168-        7667 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 5, isoform y 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 7667)
  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 7667)
  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.y-AP-His
FEATURES             Location/Qualifiers
     source          1..7667
                     /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             4247..4264
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4290..4411
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4327..4346)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4381..4400
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4449..4466)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4620..5208)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4700..4719)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5379..6170,6171..6239))
                     /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     6002..6021
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6240..6344)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6412..6430
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6468..6490)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6492..6512)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6497..6827
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6678..6813
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6740..6759
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6964..6983
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7050..7505)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7187..7208)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7399..7418
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7631..7653
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7645..7662
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7646..7662
                     /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 caggccccaa gaccaggata gtggccagct acctccgtgt
     1681 ggagttcggg ggaaaagcag ggctggtgac cactggagta agaggccgct gccagtgcca
     1741 cggccacgct gcccgctgtg ccactcgggc ccagccccca cgctgccgct gccgccacca
     1801 caccactggc ccagggtgtg agagctgtcg cccatcccat cgagactggc cctggcgacc
     1861 tgccacaccc cagcaccctc acccgtgctt gccctgttcc tgcaatcagc atgcccgccg
     1921 ctgccgattc aactccgagc tattcaggtt gtcaggtggc cgtagtgggg gtgtgtgtga
     1981 gcggtgccgc caccacacag ctgggcggca ctgtcactac tgccagccag ggttctggag
     2041 ggacccgagc cagcccatca ctagtcacaa ggcatgcagg gcctgccagt gccacccaat
     2101 tggagcaacg ggagggatgt gcaaccagac cagtggccag tgctcctgca agttaggggt
     2161 cacaggcctg acatgcaatc gctgtggtcc tggctaccag cagagccgct cacccaggat
     2221 gccctgccaa cgaattccag aggcaacaac tacccctgct actacccctg ttgcatctcg
     2281 atcagaccca cagtgtcaag gctattgcaa tgtctccgtc agcagtgtgc acatgagcct
     2341 tcagagatac tgtcagcagg attatgttct gcacgcccag gtttcagcgt cgtcgtcgca
     2401 gccgtctgag gcggtgggcc ccgagtggtg gcggctggcc gtgcacgtgc tggctgtgtt
     2461 caaacagcgt gcgtggcccg tgcgccgcgg tggccaggag gcctgggtgc cacgtgctga
     2521 cctaatctgc ggctgcctgc gcctgcgccc tggcgctgac tacctcctgc tgggccgtgc
     2581 tgcgcagacc cacgacgacg acaactatga ccctgcgcgc ctcatcctga accgccacgg
     2641 cctcgcgctg ccctggagac cacgatgggc ccggccgctg cggcgactgc agcagaagga
     2701 gcgcggaggt gcctgccgag gcctgctgcc acccacacgg agccctgggc ccaggaacgg
     2761 cgcgcctgga tcttccggaa tcatcccagt tgaggaggag aacccggact tctggaaccg
     2821 cgaggcagcc gaggccctgg gtgccgccaa gaagctgcag cctgcacaga cagccgccaa
     2881 gaacctcatc atcttcctgg gcgatgggat gggggtgtct acggtgacag ctgccaggat
     2941 cctaaaaggg cagaagaagg acaaactggg gcctgagata cccctggcca tggaccgctt
     3001 cccatatgtg gctctgtcca agacatacaa tgtagacaaa catgtgccag acagtggagc
     3061 cacagccacg gcctacctgt gcggggtcaa gggcaacttc cagaccattg gcttgagtgc
     3121 agccgcccgc tttaaccagt gcaacacgac acgcggcaac gaggtcatct ccgtgatgaa
     3181 tcgggccaag aaagcaggga agtcagtggg agtggtaacc accacacgag tgcagcacgc
     3241 ctcgccagcc ggcacctacg cccacacggt gaaccgcaac tggtactcgg acgccgacgt
     3301 gcctgcctcg gcccgccagg aggggtgcca ggacatcgct acgcagctca tctccaacat
     3361 ggacattgac gtgatcctag gtgggggccg aaagtacatg tttcgcatgg gaaccccaga
     3421 ccctgagtac ccagatgact acagccaagg tgggaccagg ctggacggga agaatctggt
     3481 gcaggaatgg ctggcgaagc gccagggtgc ccggtacgtg tggaaccgca ctgagctcat
     3541 gcaggcttcc ctggacccgt ctgtggccca tctcatgggt ctctttgagc ctggagacat
     3601 gaaatacgag atccaccgag actccacact ggacccctcc ctgatggaga tgacagaggc
     3661 tgccctgcgc ctgctgagca ggaacccccg cggcttcttc ctcttcgtgg agggtggtcg
     3721 catcgaccat ggtcatcatg aaagcagggc ttaccgggca ctgactgaga cgatcatgtt
     3781 cgacgacgcc attgagaggg cgggccagct caccagcgag gaggacacgc tgagcctcgt
     3841 cactgccgac cactcccacg tcttctcctt cggaggctac cccctgcgag ggagctccat
     3901 cttcgggctg gcccctggca aggcccggga caggaaggcc tacacggtcc tcctatacgg
     3961 aaacggtcca ggctatgtgc tcaaggacgg cgcccggccg gatgttaccg agagcgagag
     4021 cgggagcccc gagtatcggc agcagtcagc agtgcccctg gacgaagaga cccacgcagg
     4081 cgaggacgtg gcggtgttcg cgcgcggccc gcaggcgcac ctggttcacg gcgtgcagga
     4141 gcagaccttc atagcgcacg tcatggcctt cgccgcctgc ctggagccct acaccgcctg
     4201 cgacctggcg ccccccgccg gcaccaccga cgccgcgcac ccgggtcatc atcaccatca
     4261 ccattaatct agagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca
     4321 aataaagcaa tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt
     4381 gtggtttgtc caaactcatc aatgtatctt atcatgtctg gtcgactcta gctagactct
     4441 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     4501 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     4561 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     4621 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     4681 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     4741 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     4801 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     4861 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     4921 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     4981 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     5041 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     5101 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     5161 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     5221 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     5281 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     5341 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     5401 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     5461 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     5521 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     5581 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     5641 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     5701 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     5761 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     5821 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     5881 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     5941 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     6001 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     6061 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     6121 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     6181 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     6241 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     6301 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     6361 gtgccacctg acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt
     6421 atcacgaggc cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg
     6481 cagctcccgg aggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag
     6541 gcagaagtat gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag
     6601 gctccccagc aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc
     6661 cgcccctaac tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc
     6721 atggctgact aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat
     6781 tccagaagta gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag
     6841 cttgtatatc cattttcgtc ccggagacgg tcacagcttg tctgtaagcg gatgccggga
     6901 gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact
     6961 atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca
     7021 gatgcgtaag gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt
     7081 cgcgttaaat ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat
     7141 cccttataaa tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa
     7201 gagtccacta ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg
     7261 cgatggccca ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa
     7321 agcactaaat cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc
     7381 gaacgtggcg agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag
     7441 tgtagcggtc acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg
     7501 cgcgtccatt cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct
     7561 tcgctattac gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg
     7621 ccagggtttt cccagtcacg acgttgtaaa acgacggcca gtgaatt
//
