LOCUS       sequence-131290-        7537 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the entire Netrin 1 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 7537)
  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 7537)
  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-Fc-His
FEATURES             Location/Qualifiers
     source          1..7537
                     /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             4117..4134
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4160..4281
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4197..4216)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4251..4270
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4319..4336)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4490..5078)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4570..4589)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5249..6040,6041..6109))
                     /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     5872..5891
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6110..6214)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6282..6300
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6338..6360)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6362..6382)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6367..6697
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6548..6683
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6610..6629
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     6834..6853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(6920..7375)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7057..7078)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7269..7288
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7501..7523
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7515..7532
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7516..7532
                     /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 acaaaactca cacatgccca ccgtgcccag cacctgaact cctgggggga
     3481 ccgtcagtct tcctcttccc cccaaaaccc aaggacaccc tcatgatctc ccggacccct
     3541 gaggtcacat gcgtggtggt ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg
     3601 tacgtggacg gcgtggaggt gcataatgcc aagacaaagc cgcgggagga gcagtacaac
     3661 agcacgtacc gtgtggtcag cgtcctcacc gtcctgcacc aggactggct gaatggcaag
     3721 gagtacaagt gcaaggtctc caacaaagcc ctcccagccc ccatcgagaa aaccatctcc
     3781 aaagccaaag ggcagccccg agaaccacag gtgtataccc tgcccccatc ccgggatgag
     3841 ctgaccaaga accaggtcag cctgacctgc ctggtcaaag gcttctatcc cagcgacatc
     3901 gccgtggagt gggagagcaa tgggcagccg gagaacaact acaagaccac gcctcccgtg
     3961 ctggactccg acggctcctt cttcctctac agcaagctca ccgtggacaa gagcaggtgg
     4021 cagcagggga acgtcttctc atgctccgtg atgcatgagg ctctgcacaa ccactacacg
     4081 cagaagagcc tctccctgtc tccgggtaaa ggcggccatc atcaccatca ccattaatct
     4141 agagatctta ttaaagcaga acttgtttat tgcagcttat aatggttaca aataaagcaa
     4201 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     4261 caaactcatc aatgtatctt atcatgtctg gtcgactcta gctagactct tccgcttcct
     4321 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4381 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4441 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4501 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4561 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4621 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4681 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4741 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4801 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4861 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4921 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4981 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     5041 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5101 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5161 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5221 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5281 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5341 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5401 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5461 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5521 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5581 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5641 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5701 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5761 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5821 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5881 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5941 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     6001 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6061 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6121 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6181 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6241 acgtctaaga aaccattatt atcatgacat taacctataa aaataggcgt atcacgaggc
     6301 cctttcgtct cgcgcgtttc ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg
     6361 aggaatgtgt gtcagttagg gtgtggaaag tccccaggct ccccaggcag gcagaagtat
     6421 gcaaagcatg catctcaatt agtcagcaac caggtgtgga aagtccccag gctccccagc
     6481 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accatagtcc cgcccctaac
     6541 tccgcccatc ccgcccctaa ctccgcccag ttccgcccat tctccgcccc atggctgact
     6601 aatttttttt atttatgcag aggccgaggc cgcctctgcc tctgagctat tccagaagta
     6661 gtgaggaggc ttttttggag gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc
     6721 cattttcgtc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     6781 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     6841 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     6901 gagaaaatac cgcatcagga aattgtaagc gttaatattt tgttaaaatt cgcgttaaat
     6961 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     7021 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     7081 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     7141 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     7201 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     7261 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     7321 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt
     7381 cgccattcag gctgcgcaac tgttgggaag ggcgatcggt gcgggcctct tcgctattac
     7441 gccagctggc gaaaggggga tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt
     7501 cccagtcacg acgttgtaaa acgacggcca gtgaatt
//
