LOCUS       sequence-135464-        9053 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Neuropilin 2, isoform 4 
            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 9053)
  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 9053)
  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: Nrp2.4-AP-His
FEATURES             Location/Qualifiers
     source          1..9053
                     /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             5633..5650
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5676..5797
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5713..5732)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5767..5786
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5835..5852)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6006..6594)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6086..6105)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6765..7556,7557..7625))
                     /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     7388..7407
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7626..7730)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7798..7816
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7854..7876)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7878..7898)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7883..8213
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8064..8199
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8126..8145
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8350..8369
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8436..8891)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8573..8594)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8785..8804
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     9017..9039
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     9031..9048
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     9032..9048
                     /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 catgaccctc 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 gcagatgaat atgaaggaga ttggagcaac tcttcttcct ctacctcagg
     4081 ggctggtgac ccctcatctg gcaaagaaaa gagctggctg tacaccctag atcccattct
     4141 gatcggcgcg cctggatctt ccggaatcat cccagttgag gaggagaacc cggacttctg
     4201 gaaccgcgag gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc
     4261 cgccaagaac ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc
     4321 caggatccta aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga
     4381 ccgcttccca tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag
     4441 tggagccaca gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt
     4501 gagtgcagcc gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt
     4561 gatgaatcgg gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca
     4621 gcacgcctcg ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc
     4681 cgacgtgcct gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc
     4741 caacatggac attgacgtga tcctaggtgg gggccgaaag tacatgtttc gcatgggaac
     4801 cccagaccct gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa
     4861 tctggtgcag gaatggctgg cgaagcgcca gggtgcccgg tacgtgtgga accgcactga
     4921 gctcatgcag gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg
     4981 agacatgaaa tacgagatcc accgagactc cacactggac ccctccctga tggagatgac
     5041 agaggctgcc ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg
     5101 tggtcgcatc gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat
     5161 catgttcgac gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag
     5221 cctcgtcact gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag
     5281 ctccatcttc gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct
     5341 atacggaaac ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag
     5401 cgagagcggg agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca
     5461 cgcaggcgag gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt
     5521 gcaggagcag accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac
     5581 cgcctgcgac ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtcatcatca
     5641 ccatcaccat taatctagag atcttattaa agcagaactt gtttattgca gcttataatg
     5701 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     5761 ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctggtcg actctagcta
     5821 gactcttccg cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg
     5881 gtatcagctc actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga
     5941 aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg
     6001 gcgtttttcc ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag
     6061 aggtggcgaa acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc
     6121 gtgcgctctc ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg
     6181 ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt
     6241 cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc
     6301 ggtaactatc gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc
     6361 actggtaaca ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg
     6421 tggcctaact acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca
     6481 gttaccttcg gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc
     6541 ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat
     6601 cctttgatct tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt
     6661 ttggtcatga gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt
     6721 tttaaatcaa tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc
     6781 agtgaggcac ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc
     6841 gtcgtgtaga taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata
     6901 ccgcgagacc cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg
     6961 gccgagcgca gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc
     7021 cgggaagcta gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct
     7081 acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa
     7141 cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt
     7201 cctccgatcg ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca
     7261 ctgcataatt ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac
     7321 tcaaccaagt cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca
     7381 atacgggata ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt
     7441 tcttcggggc gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc
     7501 actcgtgcac ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca
     7561 aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata
     7621 ctcatactct tcctttttca atattattga agcatttatc agggttattg tctcatgagc
     7681 ggatacatat ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc
     7741 cgaaaagtgc cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat
     7801 aggcgtatca cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga
     7861 cacatgcagc tcccggagga atgtgtgtca gttagggtgt ggaaagtccc caggctcccc
     7921 aggcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     7981 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     8041 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     8101 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     8161 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     8221 cgggagcttg tatatccatt ttcgtcccgg agacggtcac agcttgtctg taagcggatg
     8281 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     8341 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     8401 cgcacagatg cgtaaggaga aaataccgca tcaggaaatt gtaagcgtta atattttgtt
     8461 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     8521 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     8581 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     8641 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     8701 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     8761 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     8821 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     8881 acagggcgcg tccattcgcc attcaggctg cgcaactgtt gggaagggcg atcggtgcgg
     8941 gcctcttcgc tattacgcca gctggcgaaa gggggatgtg ctgcaaggcg attaagttgg
     9001 gtaacgccag ggttttccca gtcacgacgt tgtaaaacga cggccagtga att
//
