LOCUS       sequence-135580-        9008 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the Neuropilin 2, isoform 6 
            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 9008)
  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 9008)
  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: Nrp2.6-AP-His
FEATURES             Location/Qualifiers
     source          1..9008
                     /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             5588..5605
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5631..5752
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5668..5687)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5722..5741
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5790..5807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5961..6549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6041..6060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6720..7511,7512..7580))
                     /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     7343..7362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7581..7685)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7753..7771
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7809..7831)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7833..7853)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7838..8168
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      8019..8154
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     8081..8100
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     8305..8324
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8391..8846)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8528..8549)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8740..8759
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8972..8994
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8986..9003
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8987..9003
                     /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 gaaacacggg cgcattatcc tgcccagcta
     3901 tgacatggag tatcagatcg tgttcgaggg agtgataggg aagggacgat cgggagagat
     3961 ttccatcgat gacattcgga taagcactga tgtcccactg gagaactgca tggaacccat
     4021 atcagctttt gcagggggca ccctcccgcc agggaccgag cccacagtgg acacggtgcc
     4081 cgtgcagccc atcccagccg gcgcgcctgg atcttccgga atcatcccag ttgaggagga
     4141 gaacccggac ttctggaacc gcgaggcagc cgaggccctg ggtgccgcca agaagctgca
     4201 gcctgcacag acagccgcca agaacctcat catcttcctg ggcgatggga tgggggtgtc
     4261 tacggtgaca gctgccagga tcctaaaagg gcagaagaag gacaaactgg ggcctgagat
     4321 acccctggcc atggaccgct tcccatatgt ggctctgtcc aagacataca atgtagacaa
     4381 acatgtgcca gacagtggag ccacagccac ggcctacctg tgcggggtca agggcaactt
     4441 ccagaccatt ggcttgagtg cagccgcccg ctttaaccag tgcaacacga cacgcggcaa
     4501 cgaggtcatc tccgtgatga atcgggccaa gaaagcaggg aagtcagtgg gagtggtaac
     4561 caccacacga gtgcagcacg cctcgccagc cggcacctac gcccacacgg tgaaccgcaa
     4621 ctggtactcg gacgccgacg tgcctgcctc ggcccgccag gaggggtgcc aggacatcgc
     4681 tacgcagctc atctccaaca tggacattga cgtgatccta ggtgggggcc gaaagtacat
     4741 gtttcgcatg ggaaccccag accctgagta cccagatgac tacagccaag gtgggaccag
     4801 gctggacggg aagaatctgg tgcaggaatg gctggcgaag cgccagggtg cccggtatgt
     4861 gtggaaccgc actgagctca tgcaggcttc cctggacccg tctgtgaccc atctcatggg
     4921 tctctttgag cctggagaca tgaaatacga gatccaccga gactccacac tggacccctc
     4981 cctgatggag atgacagagg ctgccctgcg cctgctgagc aggaaccccc gcggcttctt
     5041 cctcttcgtg gagggtggtc gcatcgacca tggtcatcat gaaagcaggg cttaccgggc
     5101 actgactgag acgatcatgt tcgacgacgc cattgagagg gcgggccagc tcaccagcga
     5161 ggaggacacg ctgagcctcg tcactgccga ccactcccac gtcttctcct tcggaggcta
     5221 ccccctgcga gggagctcca tcttcgggct ggcccctggc aaggcccggg acaggaaggc
     5281 ctacacggtc ctcctatacg gaaacggtcc aggctatgtg ctcaaggacg gcgcccggcc
     5341 ggatgttacc gagagcgaga gcgggagccc cgagtatcgg cagcagtcag cagtgcccct
     5401 ggacgaagag acccacgcag gcgaggacgt ggcggtgttc gcgcgcggcc cgcaggcgca
     5461 cctggttcac ggcgtgcagg agcagacctt catagcgcac gtcatggcct tcgccgcctg
     5521 cctggagccc tacaccgcct gcgacctggc gccccccgcc ggcaccaccg acgccgcgca
     5581 cccgggtcat catcaccatc accattaatc tagagatctt attaaagcag aacttgttta
     5641 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5701 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5761 ggtcgactct agctagactc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     5821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     5881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     5941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     6001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     6061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     6121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     6181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     6241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     6301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     6361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     6421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     6481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     6541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     6601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     6661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     6721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     6781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     6841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     6901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     6961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     7021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     7081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     7141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     7201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     7261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     7321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     7381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     7441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     7501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     7561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     7621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     7681 ccgcgcacat ttccccgaaa agtgccacct gacgtctaag aaaccattat tatcatgaca
     7741 ttaacctata aaaataggcg tatcacgagg ccctttcgtc tcgcgcgttt cggtgatgac
     7801 ggtgaaaacc tctgacacat gcagctcccg gaggaatgtg tgtcagttag ggtgtggaaa
     7861 gtccccaggc tccccaggca ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa
     7921 ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     7981 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     8041 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     8101 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     8161 tttgcaaaaa gctcccggga gcttgtatat ccattttcgt cccggagacg gtcacagctt
     8221 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     8281 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     8341 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg aaattgtaag
     8401 cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa atcagctcat tttttaacca
     8461 ataggccgaa atcggcaaaa tcccttataa atcaaaagaa tagaccgaga tagggttgag
     8521 tgttgttcca gtttggaaca agagtccact attaaagaac gtggactcca acgtcaaagg
     8581 gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa ccatcaccct aatcaagttt
     8641 tttggggtcg aggtgccgta aagcactaaa tcggaaccct aaagggagcc cccgatttag
     8701 agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa gggaagaaag cgaaaggagc
     8761 gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc gtaaccacca cacccgccgc
     8821 gcttaatgcg ccgctacagg gcgcgtccat tcgccattca ggctgcgcaa ctgttgggaa
     8881 gggcgatcgg tgcgggcctc ttcgctatta cgccagctgg cgaaaggggg atgtgctgca
     8941 aggcgattaa gttgggtaac gccagggttt tcccagtcac gacgttgtaa aacgacggcc
     9001 agtgaatt
//
