LOCUS       sequence-135549-        8616 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses the extracellular region of the NCAM1, isoform 0.0.0 
            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 8616)
  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 8616)
  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_0.0.0-AP-His
FEATURES             Location/Qualifiers
     source          1..8616
                     /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             1648..1659
                     /codon_start=1
                     /product="WELQut protease recognition and cleavage site"
                     /label=WELQut site
                     /translation="WELQ"
     CDS             5196..5213
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    5239..5360
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5276..5295)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5330..5349
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5398..5415)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5569..6157)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5649..5668)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6328..7119,7120..7188))
                     /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     6951..6970
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7189..7293)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     7361..7379
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(7417..7439)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(7441..7461)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7446..7776
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7627..7762
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7689..7708
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7913..7932
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7999..8454)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8136..8157)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     8348..8367
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8580..8602
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     8594..8611
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     8595..8611
                     /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 cgccaccatg ctgcgaacta
     1621 aggatctcat ctggactttg tttttcctgg gaactgcagt ttccctgcag gtagatattg
     1681 ttcccagcca aggagaaatc agcgttggag agtccaaatt cttcctgtgt caagtggcag
     1741 gagatgccaa agataaggac atctcctggt tctcccccaa tggggagaag ctgagcccaa
     1801 accagcagcg gatctcagtg gtgtggaatg atgacgactc ctctaccctc accatctaca
     1861 acgccaacat cgacgatgcc ggcatataca agtgtgtggt cacggctgag gacggcaccc
     1921 agtctgaggc cactgtcaac gtgaagatct tccagaagct catgttcaag aatgcaccaa
     1981 ccccacagga gtttaaggaa ggggaggatg ctgtgattgt ctgtgatgtg gtcagctccc
     2041 tgcctccaac catcatctgg aaacacaaag gccgagatgt cattctgaaa aaagacgtcc
     2101 ggttcatagt cctgtccaac aactacctgc agatcagggg catcaagaaa acagatgagg
     2161 gtacttaccg ctgtgagggc aggatcctgg cccgcgggga aatcaacttc aaggacattc
     2221 aggtcattgt gaatgtacca cccactgtcc aggccagaca gagcatcgtg aatgccactg
     2281 ccaacctggg ccagtctgtc accctggtgt gtgatgccga tggcttccca gagcccacca
     2341 tgagctggac aaaggatggg gaacccatag agaacgagga ggaagatgac gagaagcaca
     2401 tcttcagtga tgacagctcc gagctgacca tcaggaatgt ggataaaaac gacgaggccg
     2461 aatacgtctg catcgcagag aacaaggctg gcgagcagga tgcctccatc cacctcaagg
     2521 tctttgcaaa gcccaaaatc acctatgtag agaatcagac ggccatggaa ctagaggagc
     2581 aagtcactct gacctgtgaa gcctccggag accccatccc ctccatcacc tggagaacat
     2641 ccacccggaa catcagcagt gaagaaaaga ctctggatgg gcacatggtg gttcgcagcc
     2701 atgctcgtgt gtcctccttg accctgaaga gcatccagta cacagatgct ggagaataca
     2761 tctgcactgc cagcaacacc atcggccagg attcccagtc catgtacctc gaagttcaat
     2821 atgctcccaa gctccagggc cctgtggctg tgtacacttg ggaagggaac caggtgaaca
     2881 tcacctgtga ggtcttcgcc tacccaagtg ccacaatctc ctggttccga gatggtcagt
     2941 tgctgccaag ctccaactac agcaatatca agatctacaa caccccatct gctagctatc
     3001 tggaggtgac ccctgattca gaaaatgact ttggaaacta caactgtaca gcagtgaacc
     3061 gtattggaca ggagtccttg gaattcatcc ttgttcaagc agacacaccg tcttctccat
     3121 ccattgaccg ggtggaacca tactccagca cagcgcaggt gcagtttgat gagccagagg
     3181 ccacaggggg cgtgcccatt ctcaagtaca aggctgagtg gaagtcgctg ggcgaagaat
     3241 cctggcattt caagtggtat gatgccaaag aagccaacat ggaaggcatt gtcaccatca
     3301 tgggcctgaa acctgagacg aggtactcgg tacgactggc ggccctcaac ggcaaggggc
     3361 tgggcgagat cagtgcagcc actgagttca agacacagcc agtccgggaa cccagtgcac
     3421 ccaagctgga agggcagatg ggagaggacg ggaactccat caaggtgaac ctgatcaagc
     3481 aggatgacgg aggctccccc atcagacact atctggtcaa gtacagagcg ctcgcctctg
     3541 agtggaaacc ggaaatcagg ctcccatccg gcagtgacca cgtcatgctc aagtccctgg
     3601 actggaacgc agagtatgaa gtctatgtgg tagctgaaaa ccagcaagga aaatccaagg
     3661 cagctcactt tgtgttcagg acctcagccc agcccacggc catcccagcc actagtggat
     3721 cttccggaat catcccagtt gaggaggaga acccggactt ctggaaccgc gaggcagccg
     3781 aggccctggg tgccgccaag aagctgcagc ctgcacagac agccgccaag aacctcatca
     3841 tcttcctggg cgatgggatg ggggtgtcta cggtgacagc tgccaggatc ctaaaagggc
     3901 agaagaagga caaactgggg cctgagatac ccctggccat ggaccgcttc ccatatgtgg
     3961 ctctgtccaa gacatacaat gtagacaaac atgtgccaga cagtggagcc acagccacgg
     4021 cctacctgtg cggggtcaag ggcaacttcc agaccattgg cttgagtgca gccgcccgct
     4081 ttaaccagtg caacacgaca cgcggcaacg aggtcatctc cgtgatgaat cgggccaaga
     4141 aagcagggaa gtcagtggga gtggtaacca ccacacgagt gcagcacgcc tcgccagccg
     4201 gcacctacgc ccacacggtg aaccgcaact ggtactcgga cgccgacgtg cctgcctcgg
     4261 cccgccagga ggggtgccag gacatcgcta cgcagctcat ctccaacatg gacattgacg
     4321 tgatcctagg tgggggccga aagtacatgt ttcgcatggg aaccccagac cctgagtacc
     4381 cagatgacta cagccaaggt gggaccaggc tggacgggaa gaatctggtg caggaatggc
     4441 tggcgaagcg ccagggtgcc cggtacgtgt ggaaccgcac tgagctcatg caggcttccc
     4501 tggacccgtc tgtgacccat ctcatgggtc tctttgagcc tggagacatg aaatacgaga
     4561 tccaccgaga ctccacactg gacccctccc tgatggagat gacagaggct gccctgcgcc
     4621 tgctgagcag gaacccccgc ggcttcttcc tcttcgtgga gggtggtcgc atcgaccatg
     4681 gtcatcatga aagcagggct taccgggcac tgactgagac gatcatgttc gacgacgcca
     4741 ttgagagggc gggccagctc accagcgagg aggacacgct gagcctcgtc actgccgacc
     4801 actcccacgt cttctccttc ggaggctacc ccctgcgagg gagctccatc ttcgggctgg
     4861 cccctggcaa ggcccgggac aggaaggcct acacggtcct cctatacgga aacggtccag
     4921 gctatgtgct caaggacggc gcccggccgg atgttaccga gagcgagagc gggagccccg
     4981 agtatcggca gcagtcagca gtgcccctgg acgaagagac ccacgcaggc gaggacgtgg
     5041 cggtgttcgc gcgcggcccg caggcgcacc tggttcacgg cgtgcaggag cagaccttca
     5101 tagcgcacgt catggccttc gccgcctgcc tggagcccta caccgcctgc gacctggcgc
     5161 cccccgccgg caccaccgac gccgcgcacc cgggtcatca tcaccatcac cattaatcta
     5221 gagatcttat taaagcagaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     5281 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     5341 aaactcatca atgtatctta tcatgtctgg tcgactctag ctagactctt ccgcttcctc
     5401 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     5461 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     5521 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     5581 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     5641 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     5701 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     5761 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     5821 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     5881 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     5941 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtggccta actacggcta
     6001 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     6061 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     6121 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     6181 ggggtctgac gctcagtgga acgaaaactc acgttaaggg attttggtca tgagattatc
     6241 aaaaaggatc ttcacctaga tccttttaaa ttaaaaatga agttttaaat caatctaaag
     6301 tatatatgag taaacttggt ctgacagtta ccaatgctta atcagtgagg cacctatctc
     6361 agcgatctgt ctatttcgtt catccatagt tgcctgactc cccgtcgtgt agataactac
     6421 gatacgggag ggcttaccat ctggccccag tgctgcaatg ataccgcgag acccacgctc
     6481 accggctcca gatttatcag caataaacca gccagccgga agggccgagc gcagaagtgg
     6541 tcctgcaact ttatccgcct ccatccagtc tattaattgt tgccgggaag ctagagtaag
     6601 tagttcgcca gttaatagtt tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc
     6661 acgctcgtcg tttggtatgg cttcattcag ctccggttcc caacgatcaa ggcgagttac
     6721 atgatccccc atgttgtgca aaaaagcggt tagctccttc ggtcctccga tcgttgtcag
     6781 aagtaagttg gccgcagtgt tatcactcat ggttatggca gcactgcata attctcttac
     6841 tgtcatgcca tccgtaagat gcttttctgt gactggtgag tactcaacca agtcattctg
     6901 agaatagtgt atgcggcgac cgagttgctc ttgcccggcg tcaatacggg ataataccgc
     6961 gccacatagc agaactttaa aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact
     7021 ctcaaggatc ttaccgctgt tgagatccag ttcgatgtaa cccactcgtg cacccaactg
     7081 atcttcagca tcttttactt tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa
     7141 tgccgcaaaa aagggaataa gggcgacacg gaaatgttga atactcatac tcttcctttt
     7201 tcaatattat tgaagcattt atcagggtta ttgtctcatg agcggataca tatttgaatg
     7261 tatttagaaa aataaacaaa taggggttcc gcgcacattt ccccgaaaag tgccacctga
     7321 cgtctaagaa accattatta tcatgacatt aacctataaa aataggcgta tcacgaggcc
     7381 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     7441 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccaggcagg cagaagtatg
     7501 caaagcatgc atctcaatta gtcagcaacc aggtgtggaa agtccccagg ctccccagca
     7561 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccatagtccc gcccctaact
     7621 ccgcccatcc cgcccctaac tccgcccagt tccgcccatt ctccgcccca tggctgacta
     7681 atttttttta tttatgcaga ggccgaggcc gcctctgcct ctgagctatt ccagaagtag
     7741 tgaggaggct tttttggagg cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc
     7801 attttcgtcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     7861 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     7921 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     7981 agaaaatacc gcatcaggaa attgtaagcg ttaatatttt gttaaaattc gcgttaaatt
     8041 tttgttaaat cagctcattt tttaaccaat aggccgaaat cggcaaaatc ccttataaat
     8101 caaaagaata gaccgagata gggttgagtg ttgttccagt ttggaacaag agtccactat
     8161 taaagaacgt ggactccaac gtcaaagggc gaaaaaccgt ctatcagggc gatggcccac
     8221 tacgtgaacc atcaccctaa tcaagttttt tggggtcgag gtgccgtaaa gcactaaatc
     8281 ggaaccctaa agggagcccc cgatttagag cttgacgggg aaagccggcg aacgtggcga
     8341 gaaaggaagg gaagaaagcg aaaggagcgg gcgctagggc gctggcaagt gtagcggtca
     8401 cgctgcgcgt aaccaccaca cccgccgcgc ttaatgcgcc gctacagggc gcgtccattc
     8461 gccattcagg ctgcgcaact gttgggaagg gcgatcggtg cgggcctctt cgctattacg
     8521 ccagctggcg aaagggggat gtgctgcaag gcgattaagt tgggtaacgc cagggttttc
     8581 ccagtcacga cgttgtaaaa cgacggccag tgaatt
//
