LOCUS       sequence-131274-        7877 bp    DNA     circular SYN 14-APR-2025
DEFINITION  Expresses the extracellular region of the NCAM1, isoform 1.18.1 
            protein, C-terminally fused to the Fc region of human IgG1 + 6X 
            histidine tag..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7877)
  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 7877)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 14, 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-Fc-His
FEATURES             Location/Qualifiers
     source          1..7877
                     /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             4457..4474
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     polyA_signal    4500..4621
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4537..4556)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4591..4610
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4659..4676)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4830..5418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4910..4929)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5589..6380,6381..6449))
                     /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     6212..6231
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6450..6554)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6622..6640
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     primer_bind     complement(6678..6700)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(6702..6722)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6707..7037
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6888..7023
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6950..6969
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     7174..7193
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     rep_origin      complement(7260..7715)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(7397..7418)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     7609..7628
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     7841..7863
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     7855..7872
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7856..7872
                     /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 acaaaactca
     3781 cacatgccca ccgtgcccag cacctgaact cctgggggga ccgtcagtct tcctcttccc
     3841 cccaaaaccc aaggacaccc tcatgatctc ccggacccct gaggtcacat gcgtggtggt
     3901 ggacgtgagc cacgaagacc ctgaggtcaa gttcaactgg tacgtggacg gcgtggaggt
     3961 gcataatgcc aagacaaagc cgcgggagga gcagtacaac agcacgtacc gtgtggtcag
     4021 cgtcctcacc gtcctgcacc aggactggct gaatggcaag gagtacaagt gcaaggtctc
     4081 caacaaagcc ctcccagccc ccatcgagaa aaccatctcc aaagccaaag ggcagccccg
     4141 agaaccacag gtgtataccc tgcccccatc ccgggatgag ctgaccaaga accaggtcag
     4201 cctgacctgc ctggtcaaag gcttctatcc cagcgacatc gccgtggagt gggagagcaa
     4261 tgggcagccg gagaacaact acaagaccac gcctcccgtg ctggactccg acggctcctt
     4321 cttcctctac agcaagctca ccgtggacaa gagcaggtgg cagcagggga acgtcttctc
     4381 atgctccgtg atgcatgagg ctctgcacaa ccactacacg cagaagagcc tctccctgtc
     4441 tccgggtaaa ggcggccatc atcaccatca ccattaatct agagatctta ttaaagcaga
     4501 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     4561 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     4621 atcatgtctg gtcgactcta gctagactct tccgcttcct cgctcactga ctcgctgcgc
     4681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag aacagtattt
     5281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5881 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6181 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6421 agggcgacac ggaaatgttg aatactcata ctcttccttt ttcaatatta ttgaagcatt
     6481 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     6541 ataggggttc cgcgcacatt tccccgaaaa gtgccacctg acgtctaaga aaccattatt
     6601 atcatgacat taacctataa aaataggcgt atcacgaggc cctttcgtct cgcgcgtttc
     6661 ggtgatgacg gtgaaaacct ctgacacatg cagctcccgg aggaatgtgt gtcagttagg
     6721 gtgtggaaag tccccaggct ccccaggcag gcagaagtat gcaaagcatg catctcaatt
     6781 agtcagcaac caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     6841 tgcatctcaa ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa
     6901 ctccgcccag ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag
     6961 aggccgaggc cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag
     7021 gcctaggctt ttgcaaaaag ctcccgggag cttgtatatc cattttcgtc ccggagacgg
     7081 tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg
     7141 gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt gtactgagag
     7201 tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac cgcatcagga
     7261 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     7321 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     7381 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     7441 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     7501 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     7561 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     7621 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     7681 acccgccgcg cttaatgcgc cgctacaggg cgcgtccatt cgccattcag gctgcgcaac
     7741 tgttgggaag ggcgatcggt gcgggcctct tcgctattac gccagctggc gaaaggggga
     7801 tgtgctgcaa ggcgattaag ttgggtaacg ccagggtttt cccagtcacg acgttgtaaa
     7861 acgacggcca gtgaatt
//
