LOCUS       sequence-131164-        8329 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 1 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 8329)
  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 8329)
  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: Ntn1-AP-His
FEATURES             Location/Qualifiers
     source          1..8329
                     /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             4909..4926
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4952..5073
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4989..5008)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5043..5062
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5111..5128)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5282..5870)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5362..5381)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6041..6832,6833..6901))
                     /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     6664..6683
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6902..7006)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7074..7092
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7130..7152)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7154..7174)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7159..7489
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7340..7475
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7402..7421
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7626..7645
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7712..8167)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7849..7870)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8061..8080
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8293..8315
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8307..8324
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8308..8324
                     /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 caaacgccac catgatgcgc
     1621 gctgtgtggg aggcgctggc ggcgctggcg gcggtggcgt gcctggtggg cgcggtgcgc
     1681 ggcgggcccg ggcttagcat gttcgccggc caggcggcgc agcctgatcc ttgctcggat
     1741 gagaatggac acccgcgccg ctgcatcccg gactttgtca acgcggcctt cggcaaggac
     1801 gtgcgcgtgt ccagcacctg cggccggccc ccggcgcgct actgcgtggt gagcgagcgt
     1861 ggtgaagagc ggctgcgctc ctgtcacctc tgcaactctt cggatcccaa gaaagcgcac
     1921 ccgcccgcct tcctcaccga cctcaataac ccgcacaacc tgacgtgctg gcagtccgag
     1981 aactacctgc agttcccgca caacgtgacg ctcactctgt cgctcggcaa gaagtttgag
     2041 gtgacctatg tgagcctgca attctgctcg ccgcggccag agtccatggc catctacaag
     2101 tccatggact acgggcgcac gtgggtgccc ttccagttct attccacgca gtgccgcaaa
     2161 atgtacaacc ggccgcaccg cgcgcctatc accaaacaga acgagcagga ggccgtgtgc
     2221 accgactcgc acaccgacat gcgcccgctc tctggcgggc tgatcgcttt cagcacgctg
     2281 gacgggcggc cctcggcgca cgacttcgac aactcgccgg tgctgcagga ctgggtcacg
     2341 gccaccgaca tccgcgtggc tttcagccgc ctgcacacgt tcggcgacga gaacgaagac
     2401 gactcggagc tggcgcgcga ctcctattac tatgcagtgt ctgacctgca ggttggcggc
     2461 cgctgcaagt gcaacggcca cgcggcgcgt tgcgtgcgcg accgagacga cagtctggtg
     2521 tgtgactgta ggcacaacac ggccggccct gaatgcgacc gttgcaagcc cttccactac
     2581 gaccggccct ggcagcgcgc cacggcccgc gaggccaacg agtgcgtggc ctgcaactgc
     2641 aacctccatg ctcggcgctg cagattcaac atggagctct ataagctatc agggcgcaag
     2701 agcgggggag tctgtctcaa ctgccgccac aacactgcgg gccgccactg ccactactgc
     2761 aaggagggct tctaccgaga catgggcaag cctatcaccc accggaaggc ttgcaaagcc
     2821 tgtgattgcc acccagtggg tgctgctggc aagacctgca atcaaaccac tggccaatgt
     2881 ccctgcaagg acggcgtgac gggcatcacc tgcaaccgat gtgccaaagg ctaccagcag
     2941 agccgctccc ccatcgcccc ttgcatcaag attcctgtgg cgccacccac cactgcagcc
     3001 agcagcgtgg aggaaccgga agactgtgac tcctattgca aggcctccaa aggcaagctg
     3061 aagatgaaca tgaagaaata ctgcaggaag gactatgctg tccagatcca catcctgaag
     3121 gccgacaaag caggggactg gtggaagttc accgtgaaca tcatctccgt gtacaagcag
     3181 ggcacaagtc gtattcgccg tggtgaccag agtttgtgga tccgctcacg agacatcgcc
     3241 tgcaagtgtc ccaaaatcaa gcccctcaag aagtacttgc tgttgggtaa tgccgaggac
     3301 tcacctgacc agagtggcat cgtggcagac aagagcagcc tggtgatcca gtggcgggac
     3361 acatgggcac ggcggctgcg caagttccag caacgggaga agaagggcaa gtgcaagaag
     3421 gccactagtg gatcttccgg aatcatccca gttgaggagg agaacccgga cttctggaac
     3481 cgcgaggcag ccgaggccct gggtgccgcc aagaagctgc agcctgcaca gacagccgcc
     3541 aagaacctca tcatcttcct gggcgatggg atgggggtgt ctacggtgac agctgccagg
     3601 atcctaaaag ggcagaagaa ggacaaactg gggcctgaga tacccctggc catggaccgc
     3661 ttcccatatg tggctctgtc caagacatac aatgtagaca aacatgtgcc agacagtgga
     3721 gccacagcca cggcctacct gtgcggggtc aagggcaact tccagaccat tggcttgagt
     3781 gcagccgccc gctttaacca gtgcaacacg acacgcggca acgaggtcat ctccgtgatg
     3841 aatcgggcca agaaagcagg gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac
     3901 gcctcgccag ccggcaccta cgcccacacg gtgaaccgca actggtactc ggacgccgac
     3961 gtgcctgcct cggcccgcca ggaggggtgc caggacatcg ctacgcagct catctccaac
     4021 atggacattg acgtgatcct aggtgggggc cgaaagtaca tgtttcgcat gggaacccca
     4081 gaccctgagt acccagatga ctacagccaa ggtgggacca ggctggacgg gaagaatctg
     4141 gtgcaggaat ggctggcgaa gcgccagggt gcccggtacg tgtggaaccg cactgagctc
     4201 atgcaggctt ccctggaccc gtctgtggcc catctcatgg gtctctttga gcctggagac
     4261 atgaaatacg agatccaccg agactccaca ctggacccct ccctgatgga gatgacagag
     4321 gctgccctgc gcctgctgag caggaacccc cgcggcttct tcctcttcgt ggagggtggt
     4381 cgcatcgacc atggtcatca tgaaagcagg gcttaccggg cactgactga gacgatcatg
     4441 ttcgacgacg ccattgagag ggcgggccag ctcaccagcg aggaggacac gctgagcctc
     4501 gtcactgccg accactccca cgtcttctcc ttcggaggct accccctgcg agggagctcc
     4561 atcttcgggc tggcccctgg caaggcccgg gacaggaagg cctacacggt cctcctatac
     4621 ggaaacggtc caggctatgt gctcaaggac ggcgcccggc cggatgttac cgagagcgag
     4681 agcgggagcc ccgagtatcg gcagcagtca gcagtgcccc tggacgaaga gacccacgca
     4741 ggcgaggacg tggcggtgtt cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag
     4801 gagcagacct tcatagcgca cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc
     4861 tgcgacctgg cgccccccgc cggcaccacc gacgccgcgc acccgggtca tcatcaccat
     4921 caccattaat ctagagatct tattaaagca gaacttgttt attgcagctt ataatggtta
     4981 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     5041 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggtcgactc tagctagact
     5101 cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat
     5161 cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga
     5221 acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt
     5281 ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt
     5341 ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc
     5401 gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa
     5461 gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct
     5521 ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta
     5581 actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg
     5641 gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc
     5701 ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta
     5761 ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg
     5821 gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt
     5881 tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg
     5941 tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta
     6001 aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg
     6061 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     6121 tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc
     6181 gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg
     6241 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     6301 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag
     6361 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     6421 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     6481 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     6541 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     6601 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     6661 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     6721 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     6781 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     6841 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     6901 tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     6961 acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa
     7021 aagtgccacc tgacgtctaa gaaaccatta ttatcatgac attaacctat aaaaataggc
     7081 gtatcacgag gccctttcgt ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca
     7141 tgcagctccc ggaggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccaggc
     7201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     7261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     7321 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     7381 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     7441 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     7501 agcttgtata tccattttcg tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     7561 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     7621 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     7681 cagatgcgta aggagaaaat accgcatcag gaaattgtaa gcgttaatat tttgttaaaa
     7741 ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa
     7801 atcccttata aatcaaaaga atagaccgag atagggttga gtgttgttcc agtttggaac
     7861 aagagtccac tattaaagaa cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag
     7921 ggcgatggcc cactacgtga accatcaccc taatcaagtt ttttggggtc gaggtgccgt
     7981 aaagcactaa atcggaaccc taaagggagc ccccgattta gagcttgacg gggaaagccg
     8041 gcgaacgtgg cgagaaagga agggaagaaa gcgaaaggag cgggcgctag ggcgctggca
     8101 agtgtagcgg tcacgctgcg cgtaaccacc acacccgccg cgcttaatgc gccgctacag
     8161 ggcgcgtcca ttcgccattc aggctgcgca actgttggga agggcgatcg gtgcgggcct
     8221 cttcgctatt acgccagctg gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa
     8281 cgccagggtt ttcccagtca cgacgttgta aaacgacggc cagtgaatt
//
