LOCUS       sequence-136369-        8669 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the NCAM1, isoform 1.18.1 
            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 8669)
  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 8669)
  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: Ncam1_1.18.1-AP-His
FEATURES             Location/Qualifiers
     source          1..8669
                     /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             1650..1661
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5249..5266
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5292..5413
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5329..5348)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5383..5402
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5451..5468)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5622..6210)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5702..5721)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6381..7172,7173..7241))
                     /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     7004..7023
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7242..7346)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7414..7432
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7470..7492)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7494..7514)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7499..7829
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7680..7815
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7742..7761
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7966..7985
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(8052..8507)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8189..8210)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8401..8420
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8633..8655
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8647..8664
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8648..8664
                     /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 tgctgcgaac
     1621 taaggatctc atctggactt tgtttttcct gggaactgca gtttccctgc aggtagatat
     1681 tgttcccagc caaggagaaa tcagcgttgg agagtccaaa ttcttcctgt gtcaagtggc
     1741 aggagatgcc aaagataagg acatctcctg gttctccccc aatggggaga agctgagccc
     1801 aaaccagcag cggatctcag tggtgtggaa tgatgacgac tcctctaccc tcaccatcta
     1861 caacgccaac atcgacgatg ccggcatata caagtgtgtg gtcacggctg aggacggcac
     1921 ccagtctgag gccactgtca acgtgaagat cttccagaag ctcatgttca agaatgcacc
     1981 aaccccacag gagtttaagg aaggggagga tgctgtgatt gtctgtgatg tggtcagctc
     2041 cctgcctcca accatcatct ggaaacacaa aggccgagat gtcattctga aaaaagacgt
     2101 ccggttcata gtcctgtcca acaactacct gcagatcagg ggcatcaaga aaacagatga
     2161 gggtacttac cgctgtgagg gcaggatcct ggcccgcggg gaaatcaact tcaaggacat
     2221 tcaggtcatt gtgaatgtac cacccactgt ccaggccaga cagagcatcg tgaatgccac
     2281 tgccaacctg ggccagtctg tcaccctggt gtgtgatgcc gatggcttcc cagagcccac
     2341 catgagctgg acaaaggatg gggaacccat agagaacgag gaggaagatg acgagaagca
     2401 catcttcagt gatgacagct ccgagctgac catcaggaat gtggataaaa acgacgaggc
     2461 cgaatacgtc tgcatcgcag agaacaaggc tggcgagcag gatgcctcca tccacctcaa
     2521 ggtctttgca aagcccaaaa tcacctatgt agagaatcag acggccatgg aactagagga
     2581 gcaagtcact ctgacctgtg aagcctccgg agaccccatc ccctccatca cctggagaac
     2641 atccacccgg aacatcagca gtgaagaaaa ggcatcgtgg actcgaccag agaagcaaga
     2701 gactctggat gggcacatgg tggttcgcag ccatgctcgt gtgtcctcct tgaccctgaa
     2761 gagcatccag tacacagatg ctggagaata catctgcact gccagcaaca ccatcggcca
     2821 ggattcccag tccatgtacc tcgaagttca atatgctccc aagctccagg gccctgtggc
     2881 tgtgtacact tgggaaggga accaggtgaa catcacctgt gaggtcttcg cctacccaag
     2941 tgccacaatc tcctggttcc gagatggtca gttgctgcca agctccaact acagcaatat
     3001 caagatctac aacaccccat ctgctagcta tctggaggtg acccctgatt cagaaaatga
     3061 ctttggaaac tacaactgta cagcagtgaa ccgtattgga caggagtcct tggaattcat
     3121 ccttgttcaa gcagacacac cgtcttctcc atccattgac cgggtggaac catactccag
     3181 cacagcgcag gtgcagtttg atgagccaga ggccacaggg ggcgtgccca ttctcaagta
     3241 caaggctgag tggaagtcgc tgggcgaaga atcctggcat ttcaagtggt atgatgccaa
     3301 agaagccaac atggaaggca ttgtcaccat catgggcctg aaacctgaga cgaggtactc
     3361 ggtacgactg gcggccctca acggcaaggg gctgggcgag atcagtgcag ccactgagtt
     3421 caagacacag ccagtccata gccctcctcc acaaggggaa cccagtgcac ccaagctgga
     3481 agggcagatg ggagaggacg ggaactccat caaggtgaac ctgatcaagc aggatgacgg
     3541 aggctccccc atcagacact atctggtcaa gtacagagcg aagctcgcct ctgagtggaa
     3601 accggaaatc aggctcccat ccggcagtga ccacgtcatg ctcaagtccc tggactggaa
     3661 cgcagagtat gaagtctatg tggtagctga aaaccagcaa ggaaaatcca aggcagctca
     3721 ctttgtgttc aggacctcag cccagcccac ggccatccca gccactagtg gatcttccgg
     3781 aatcatccca gttgaggagg agaacccgga cttctggaac cgcgaggcag ccgaggccct
     3841 gggtgccgcc aagaagctgc agcctgcaca gacagccgcc aagaacctca tcatcttcct
     3901 gggcgatggg atgggggtgt ctacggtgac agctgccagg atcctaaaag ggcagaagaa
     3961 ggacaaactg gggcctgaga tacccctggc catggaccgc ttcccatatg tggctctgtc
     4021 caagacatac aatgtagaca aacatgtgcc agacagtgga gccacagcca cggcctacct
     4081 gtgcggggtc aagggcaact tccagaccat tggcttgagt gcagccgccc gctttaacca
     4141 gtgcaacacg acacgcggca acgaggtcat ctccgtgatg aatcgggcca agaaagcagg
     4201 gaagtcagtg ggagtggtaa ccaccacacg agtgcagcac gcctcgccag ccggcaccta
     4261 cgcccacacg gtgaaccgca actggtactc ggacgccgac gtgcctgcct cggcccgcca
     4321 ggaggggtgc caggacatcg ctacgcagct catctccaac atggacattg acgtgatcct
     4381 aggtgggggc cgaaagtaca tgtttcgcat gggaacccca gaccctgagt acccagatga
     4441 ctacagccaa ggtgggacca ggctggacgg gaagaatctg gtgcaggaat ggctggcgaa
     4501 gcgccagggt gcccggtatg tgtggaaccg cactgagctc atgcaggctt ccctggaccc
     4561 gtctgtgacc catctcatgg gtctctttga gcctggagac atgaaatacg agatccaccg
     4621 agactccaca ctggacccct ccctgatgga gatgacagag gctgccctgc gcctgctgag
     4681 caggaacccc cgcggcttct tcctcttcgt ggagggtggt cgcatcgacc atggtcatca
     4741 tgaaagcagg gcttaccggg cactgactga gacgatcatg ttcgacgacg ccattgagag
     4801 ggcgggccag ctcaccagcg aggaggacac gctgagcctc gtcactgccg accactccca
     4861 cgtcttctcc ttcggaggct accccctgcg agggagctcc atcttcgggc tggcccctgg
     4921 caaggcccgg gacaggaagg cctacacggt cctcctatac ggaaacggtc caggctatgt
     4981 gctcaaggac ggcgcccggc cggatgttac cgagagcgag agcgggagcc ccgagtatcg
     5041 gcagcagtca gcagtgcccc tggacgaaga gacccacgca ggcgaggacg tggcggtgtt
     5101 cgcgcgcggc ccgcaggcgc acctggttca cggcgtgcag gagcagacct tcatagcgca
     5161 cgtcatggcc ttcgccgcct gcctggagcc ctacaccgcc tgcgacctgg cgccccccgc
     5221 cggcaccacc gacgccgcgc acccgggtca tcatcaccat caccattaat ctagagatct
     5281 tattaaagca gaacttgttt attgcagctt ataatggtta caaataaagc aatagcatca
     5341 caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg tccaaactca
     5401 tcaatgtatc ttatcatgtc tggtcgactc tagctagact cttccgcttc ctcgctcact
     5461 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     5521 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     5581 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     5641 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     5701 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     5761 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     5821 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     5881 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     5941 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     6001 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     6061 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     6121 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     6181 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     6241 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     6301 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     6361 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     6421 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     6481 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     6541 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     6601 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     6661 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     6721 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     6781 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     6841 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     6901 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     6961 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     7021 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     7081 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     7141 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     7201 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     7261 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     7321 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgacgtctaa
     7381 gaaaccatta ttatcatgac attaacctat aaaaataggc gtatcacgag gccctttcgt
     7441 ctcgcgcgtt tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggaggaatgt
     7501 gtgtcagtta gggtgtggaa agtccccagg ctccccaggc aggcagaagt atgcaaagca
     7561 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     7621 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     7681 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     7741 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     7801 gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg
     7861 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     7921 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     7981 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     8041 accgcatcag gaaattgtaa gcgttaatat tttgttaaaa ttcgcgttaa atttttgtta
     8101 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
     8161 atagaccgag atagggttga gtgttgttcc agtttggaac aagagtccac tattaaagaa
     8221 cgtggactcc aacgtcaaag ggcgaaaaac cgtctatcag ggcgatggcc cactacgtga
     8281 accatcaccc taatcaagtt ttttggggtc gaggtgccgt aaagcactaa atcggaaccc
     8341 taaagggagc ccccgattta gagcttgacg gggaaagccg gcgaacgtgg cgagaaagga
     8401 agggaagaaa gcgaaaggag cgggcgctag ggcgctggca agtgtagcgg tcacgctgcg
     8461 cgtaaccacc acacccgccg cgcttaatgc gccgctacag ggcgcgtcca ttcgccattc
     8521 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
     8581 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
     8641 cgacgttgta aaacgacggc cagtgaatt
//
