LOCUS       sequence-131162-        9023 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Neuropilin 2, isoform 5 
            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 9023)
  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 9023)
  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: Nrp2.5-AP-His
FEATURES             Location/Qualifiers
     source          1..9023
                     /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)"
     regulatory      1604..1613
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             5603..5620
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5646..5767
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5683..5702)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5737..5756
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5805..5822)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5976..6564)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6056..6075)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6735..7526,7527..7595))
                     /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     7358..7377
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7596..7700)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7768..7786
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7824..7846)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7848..7868)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7853..8183
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8034..8169
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8096..8115
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8320..8339
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8406..8861)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8543..8564)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8755..8774
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8987..9009
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9001..9018
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9002..9018
                     /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 tggatatgtt
     1621 tcctcttacc tgggttttct tagctctgta cttttcagga cacgaagtga gaagccagca
     1681 agatccaccc tgcggaggtc ggctgaattc caaagatgct ggctacatca cttccccagg
     1741 ctacccccag gactatccct cccaccagaa ctgtgagtgg attgtctacg cccccgaacc
     1801 caaccagaag attgttctca acttcaaccc tcactttgaa atcgagaaac acgactgcaa
     1861 gtatgacttc attgagattc gggatgggga cagtgagtca gctgacctcc tgggcaagca
     1921 ctgtgggaac atcgccccgc ccaccatcat ctcctcaggc tccgtgttat acatcaagtt
     1981 cacctcagac tacgcccggc agggggcagg tttctctcta cgctatgaga tcttcaaaac
     2041 aggctctgaa gattgttcca agaactttac aagccccaat gggaccattg aatctccagg
     2101 gtttccagag aagtatccac acaatctgga ctgtaccttc accatcctgg ccaaacccag
     2161 gatggagatc atcctacagt tcctgacctt tgacctggag catgaccccc tacaagtggg
     2221 ggaaggagac tgtaaatatg actggctgga catctgggat ggcattccac atgttggacc
     2281 tctgattggc aagtactgtg ggacgaaaac accctccaaa ctccgctcgt ccacggggat
     2341 cctctccttg acctttcaca cggacatggc agtggccaag gatggcttct ccgcacgtta
     2401 ctatttgatc caccaggagc cacctgagaa ttttcagtgc aatgtccctt tgggaatgga
     2461 gtctggccgg attgctaatg aacagatcag tgcctcctcc accttctctg atgggaggtg
     2521 gactcctcaa cagagccggc tccatggtga tgacaatggc tggacaccca atttggattc
     2581 caacaaggag tatctccagg tggacctgcg cttcctaacc atgctcacag ccattgcaac
     2641 acagggagcc atttccaggg aaacccagaa aggctactac gtcaaatcgt acaagctgga
     2701 agtcagcaca aatggtgaag attggatggt ctaccggcat ggcaaaaacc acaagatatt
     2761 ccaagcgaac aatgatgcga ccgaggtggt gctaaacaag ctccacatgc cactgctgac
     2821 tcggttcatc aggatccgcc cgcagacgtg gcatttgggc attgcccttc gcctggagct
     2881 ctttggctgc cgggtcacag atgcaccctg ctccaacatg ctggggatgc tctcgggcct
     2941 cattgctgat acccagatct ctgcctcctc cacccgagag tacctctgga gccccagtgc
     3001 tgcccgcctg gttagtagcc gctctggctg gtttcctcgg aaccctcaag cccagccagg
     3061 tgaagaatgg cttcaggtag acctggggac acccaagaca gtgaaagggg tcatcatcca
     3121 gggagcccga ggaggagaca gcatcactgc cgtggaagcc agggcgtttg tacgcaagtt
     3181 caaagtctcc tacagcctaa atggcaagga ctgggaatat atccaggacc ccaggactca
     3241 gcagacaaag ctgtttgaag ggaacatgca ctatgacacc cctgacatcc gaaggttcga
     3301 tcctgttcca gcgcagtatg tgcgggtgta cccagagagg tggtcgccag caggcatcgg
     3361 gatgaggctg gaggtgctgg gctgtgactg gacagactca aagcccacag tggagacgct
     3421 gggacccacc gtgaagagtg aagagactac caccccatat cccatggatg aggatgccac
     3481 cgagtgtggg gaaaactgca gctttgagga tgacaaagat ttgcaacttc cttcaggatt
     3541 caactgcaac tttgattttc cggaagagac ctgtggttgg gtgtacgacc atgccaagtg
     3601 gctccggagc acgtggatca gcagcgctaa ccccaatgac agaacatttc cagatgacaa
     3661 gaacttcttg aaactgcaga gtgatggccg acgagagggc cagtacgggc ggctcatcag
     3721 cccaccggtg cacctgcccc gaagccctgt gtgcatggag ttccagtacc aagccatggg
     3781 cggccacggg gtggcactgc aggtggttcg ggaagccagc caggaaagca aactcctttg
     3841 ggtcatccgt gaggaccagg gcagcgagtg gaagcacggg cgcattatcc tgcccagcta
     3901 tgacatggag tatcagatcg tgttcgaggg agtgataggg aagggacgat cgggagagat
     3961 ttccatcgat gacattcgga taagcactga tgtcccactg gagaactgca tggaacccat
     4021 atcagctttt gcaggtgagg attttaaagg gggcaccctc ccgccaggga ccgagcccac
     4081 agtggacacg gtgcccgtgc agcccatccc agccggcgcg cctggatctt ccggaatcat
     4141 cccagttgag gaggagaacc cggacttctg gaaccgcgag gcagccgagg ccctgggtgc
     4201 cgccaagaag ctgcagcctg cacagacagc cgccaagaac ctcatcatct tcctgggcga
     4261 tgggatgggg gtgtctacgg tgacagctgc caggatccta aaagggcaga agaaggacaa
     4321 actggggcct gagatacccc tggccatgga ccgcttccca tatgtggctc tgtccaagac
     4381 atacaatgta gacaaacatg tgccagacag tggagccaca gccacggcct acctgtgcgg
     4441 ggtcaagggc aacttccaga ccattggctt gagtgcagcc gcccgcttta accagtgcaa
     4501 cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg gccaagaaag cagggaagtc
     4561 agtgggagtg gtaaccacca cacgagtgca gcacgcctcg ccagccggca cctacgccca
     4621 cacggtgaac cgcaactggt actcggacgc cgacgtgcct gcctcggccc gccaggaggg
     4681 gtgccaggac atcgctacgc agctcatctc caacatggac attgacgtga tcctaggtgg
     4741 gggccgaaag tacatgtttc gcatgggaac cccagaccct gagtacccag atgactacag
     4801 ccaaggtggg accaggctgg acgggaagaa tctggtgcag gaatggctgg cgaagcgcca
     4861 gggtgcccgg tacgtgtgga accgcactga gctcatgcag gcttccctgg acccgtctgt
     4921 ggcccatctc atgggtctct ttgagcctgg agacatgaaa tacgagatcc accgagactc
     4981 cacactggac ccctccctga tggagatgac agaggctgcc ctgcgcctgc tgagcaggaa
     5041 cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc gaccatggtc atcatgaaag
     5101 cagggcttac cgggcactga ctgagacgat catgttcgac gacgccattg agagggcggg
     5161 ccagctcacc agcgaggagg acacgctgag cctcgtcact gccgaccact cccacgtctt
     5221 ctccttcgga ggctaccccc tgcgagggag ctccatcttc gggctggccc ctggcaaggc
     5281 ccgggacagg aaggcctaca cggtcctcct atacggaaac ggtccaggct atgtgctcaa
     5341 ggacggcgcc cggccggatg ttaccgagag cgagagcggg agccccgagt atcggcagca
     5401 gtcagcagtg cccctggacg aagagaccca cgcaggcgag gacgtggcgg tgttcgcgcg
     5461 cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag accttcatag cgcacgtcat
     5521 ggccttcgcc gcctgcctgg agccctacac cgcctgcgac ctggcgcccc ccgccggcac
     5581 caccgacgcc gcgcacccgg gtcatcatca ccatcaccat taatctagag atcttattaa
     5641 agcagaactt gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt
     5701 tcacaaataa agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg
     5761 tatcttatca tgtctggtcg actctagcta gactcttccg cttcctcgct cactgactcg
     5821 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     5881 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     5941 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     6001 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     6061 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     6121 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     6181 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     6241 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     6301 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     6361 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     6421 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     6481 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     6541 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     6601 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     6661 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     6721 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     6781 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     6841 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     6901 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     6961 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     7021 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     7081 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     7141 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     7201 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     7261 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     7321 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     7381 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     7441 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     7501 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     7561 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     7621 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     7681 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt ctaagaaacc
     7741 attattatca tgacattaac ctataaaaat aggcgtatca cgaggccctt tcgtctcgcg
     7801 cgtttcggtg atgacggtga aaacctctga cacatgcagc tcccggagga atgtgtgtca
     7861 gttagggtgt ggaaagtccc caggctcccc aggcaggcag aagtatgcaa agcatgcatc
     7921 tcaattagtc agcaaccagg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     7981 aaagcatgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     8041 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     8101 atgcagaggc cgaggccgcc tctgcctctg agctattcca gaagtagtga ggaggctttt
     8161 ttggaggcct aggcttttgc aaaaagctcc cgggagcttg tatatccatt ttcgtcccgg
     8221 agacggtcac agcttgtctg taagcggatg ccgggagcag acaagcccgt cagggcgcgt
     8281 cagcgggtgt tggcgggtgt cggggctggc ttaactatgc ggcatcagag cagattgtac
     8341 tgagagtgca ccatatgcgg tgtgaaatac cgcacagatg cgtaaggaga aaataccgca
     8401 tcaggaaatt gtaagcgtta atattttgtt aaaattcgcg ttaaattttt gttaaatcag
     8461 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     8521 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     8581 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     8641 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     8701 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     8761 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     8821 caccacaccc gccgcgctta atgcgccgct acagggcgcg tccattcgcc attcaggctg
     8881 cgcaactgtt gggaagggcg atcggtgcgg gcctcttcgc tattacgcca gctggcgaaa
     8941 gggggatgtg ctgcaaggcg attaagttgg gtaacgccag ggttttccca gtcacgacgt
     9001 tgtaaaacga cggccagtga att
//
