LOCUS       sequence-396955-        7666 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7666)
  AUTHORS   Wojtowicz WM, Vielmetter J, Fernandes RA, Siepe DH, Eastman CL, 
            Chisholm GB, Cox S, Klock H, Anderson PW, Rue SM, Miller JJ, Glaser 
            SM, Bragstad ML, Vance J, Lam AW, Lesley SA, Zinn K, Garcia KC
  TITLE     A Human IgSF Cell-Surface Interactome Reveals a Complex Network of 
            Protein-Protein Interactions.
  JOURNAL   Cell. 2020 Aug 20;182(4):1027-1043.e17. doi: 
            10.1016/j.cell.2020.07.025.
  PUBMED    32822567
REFERENCE   2  (bases 1 to 7666)
  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"; doi: 
            "10.1016/j.cell.2020.07"; journalName: "Cell"; date: "2020-08-20- 
            20"; volume: "182"; issue: "4"; pages: "1027-1043.e17"
COMMENT     Sequence Label: pD649-HAsp-LINGO1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7666
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             3242..3286
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             3287..3304
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3444..3461)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3450..3561
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3679..3699)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3684..4013
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3864..3999
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3926..3945
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        4061..4108
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             4127..4726
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     primer_bind     complement(4127..4146)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4623..4643
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4856..4977
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4893..4912)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4947..4966
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5030..5821,5822..5890))
                     /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     5653..5672
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5891..5982)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     6024..6043
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     6049..6068
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      6154..6742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6643..6662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6896..6913
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     7011..7028
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     7012..7028
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7484..7500
                     /label=lac operator
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="The lac repressor binds to the lac operator to 
                     inhibit transcription in E. coli. This inhibition can be 
                     relieved by adding lactose or 
                     isopropyl-beta-D-thiogalactopyranoside (IPTG)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cctgtcctcc tagatgtgaa tgtagcgccc aggacagagc
     1021 cgtgctgtgt caccggaaga gatttgtggc tgtgcccgag ggcatcccca ccgaaacaag
     1081 actgctggac ctgggcaaga accggatcaa gaccctgaac caggacgagt tcgccagctt
     1141 tccccacctg gaagaactgg aactgaacga gaacatcgtg tccgccgtgg aacctggcgc
     1201 cttcaacaac ctgttcaacc tgagaaccct gggcctgaga agcaaccggc tgaagctgat
     1261 tcctctgggc gtgttcaccg gcctgagcaa cctgaccaag ctggacatca gcgagaacaa
     1321 gatcgtgatc ctgctggact acatgttcca ggacctgtac aatctgaagt ccctggaagt
     1381 gggcgacaac gacctggtgt acatcagcca cagagccttc agcggcctga acagcctgga
     1441 acagctgacc ctggaaaagt gcaatctgac cagcattccc acagaggccc tgagccatct
     1501 gcacggactg atcgtgctga gactgcggca cctgaacatc aacgccatcc gggactacag
     1561 cttcaagcgg ctgtacagac tgaaggtgct ggaaatcagc cactggcctt acctggacac
     1621 catgacaccc aactgcctgt acggcctgaa tctgacaagc ctgagcatca cccactgtaa
     1681 cctgaccgcc gtgccttatc tggccgtcag acatctggtg tacctgcggt tcctgaacct
     1741 gagctacaac cccatcagca ccatcgaggg ctccatgctg cacgaactgc tgcggctgca
     1801 agagatccag ctcgttggag gacagctggc cgtggtggaa ccctatgcct ttcggggcct
     1861 gaactacctg agagtgctga acgtgtccgg caaccagctg accacactgg aagagtccgt
     1921 gttccacagc gtgggcaacc tggaaaccct gatcctggac agcaatcccc tggcctgcga
     1981 ttgcagactg ctgtgggtgt tccgaagaag atggcggctg aacttcaacc ggcagcagcc
     2041 tacatgtgcc acacctgagt ttgtgcaggg caaagagttc aaggacttcc ccgacgtgct
     2101 gctgcccaac tacttcacat gcagacgggc cagaatccgg gacagaaagg cccagcaggt
     2161 tttcgtggat gagggccaca cagtgcagtt tgtgtgcaga gccgatggcg atcctcctcc
     2221 tgccattctc tggctgagcc ccagaaagca cctggtgtcc gccaagagca acggcagact
     2281 gaccgtgttt cctgacggca ctctggaagt ccgctatgcc caggtgcagg acaatggcac
     2341 ctacctgtgt atcgccgcca atgccggcgg aaacgatagc atgcctgctc accttcacgt
     2401 gcggagctac agccctgatt ggcctcacca gcctaacaag accttcgcct tcatcagcaa
     2461 ccagcctggc gagggcgaag ccaatagcac cagagccaca gtgccctttc cattcgacat
     2521 caagacactg atcattgcca caggcgcgcc gggcggcgac aagacccaca cctgcccccc
     2581 ctgcccagct ccagaactgc tgggagggcc ttccgtgttc ctgttccccc ccaagcccaa
     2641 ggacaccctg atgatcagca ggacccccga ggtgacctgc gtggtggtgg ccgtgtccca
     2701 cgaggaccca gaggtgaagt tcaactggta cgtggacggc gtggaggtgc acaacgccaa
     2761 gaccaagccc agagaggagc agtacaacag cacctacagg gtggtgtccg tgctgaccgt
     2821 gctgcaccag gactggctga acggcaaaga atacaagtgc aaagtctcca acaaggccct
     2881 ggctgcccca atcgaaaaga caatcagcaa ggccaagggc cagccacggg agccccaggt
     2941 gtacaccctg ccccccagcc gggaggagat gaccaagaac caggtgtccc tgacctgtct
     3001 ggtgaagggc ttctacccca gcgatatcgc cgtggagtgg gagagcaacg gccagcccga
     3061 gaacaactac aagaccaccc ccccagtgct ggacagcgac ggcagcttct tcctgtacag
     3121 caagctgacc gtggacaagt ccaggtggca gcagggcaac gtgttcagct gcagcgtgat
     3181 gcacgaggcc ctgcacaacc actacaccca gaagtccctg agcctgagcc ccggcggcag
     3241 cggtcttaat gatattttcg aggctcagaa gattgagtgg catgaacacc accaccatca
     3301 ccactaaggc cggcctgtag tcactggcac aacgcgttac ccggtaagcc aatcgggtat
     3361 acacggtcgt catactgcag acagggttct tctactttgc aagatagtct tgagtagtaa
     3421 aataaataga tagagaaaat cagcctcgac tgtgccttct agttgccagc catctgttgt
     3481 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     3541 ataaaatgag gaaattgcat cacaacactc aaccctatct cggtctattc ttttgattta
     3601 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     3661 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3721 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3781 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3841 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3901 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3961 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     4021 cgggagcttg tatatccatt ttcggatctg atcagcacgt gttgacaatt aatcatcggc
     4081 atagtatatc ggcatagtat aatacgacaa ggtgaggaac taaaccatga ccgagtacaa
     4141 gcccacggtg cgcctcgcca cccgcgacga cgtccccagg gccgtacgca ccctcgccgc
     4201 cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat ccggaccgcc acatcgagcg
     4261 ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg ctcgacatcg gcaaggtgtg
     4321 ggtcgcggac gacggcgccg cggtggcggt ctggaccacg ccggagagcg tcgaagcggg
     4381 ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg agcggttccc ggctggccgc
     4441 gcagcaacag atggaaggcc tcctggcgcc gcaccggccc aaggagcccg cgtggttcct
     4501 ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt ctgggcagcg ccgtcgtgct
     4561 ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc ttcctggaga catccgcgcc
     4621 ccgcaacctc cccttctacg agcggctcgg cttcaccgtc accgccgacg tcgaggtgcc
     4681 cgaaggaccg cgcacctggt gcatgacccg caagcccggt gcctgacacg tgctacgaga
     4741 tttcgattcc accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc
     4801 cggctggatg atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt
     4861 gtttattgca gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa
     4921 agcatttttt tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca
     4981 tgtctgtata ccgtcgacct ctagctagag cttggcgtaa tcatggtcat taccaatgct
     5041 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     5101 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agcgctgcga
     5161 tgataccgcg agaaccacgc tcaccggctc cggatttatc agcaataaac cagccagccg
     5221 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5281 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5341 tcgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5401 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5461 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5521 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5581 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5641 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5701 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5761 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5821 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5881 gaatactcat attcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5941 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtc agtgttacaa
     6001 ccaattaacc aattctgaac attatcgcga gcccatttat acctgaatat ggctcataac
     6061 accccttgct catgaccaaa atcccttaac gtgagttacg cgcgcgtcgt tccactgagc
     6121 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     6181 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     6241 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     6301 tcttctagtg tagccgtagt tagcccacca cttcaagaac tctgtagcac cgcctacata
     6361 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     6421 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     6481 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6541 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6601 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6661 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6721 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6781 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6841 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6901 gtcagtgagc gaggaagcgg aaggcgagag tagggaactg ccaggcatca aactaagcag
     6961 aaggcccctg acggatggcc tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga
     7021 cggccagtct taagctcggg ccccctgggc ggttctgata acgagtaatc gttaatccgc
     7081 aaataacgta aaaacccgct tcggcgggtt tttttatggg gggagtttag ggaaagagca
     7141 tttgtcagaa tatttaaggg cgcctgtcac tttgcttgat atatgagaat tatttaacct
     7201 tataaatgag aaaaaagcaa cgcactttaa ataagatacg ttgctttttc gattgatgaa
     7261 cacctataat taaactattc atctattatt tatgattttt tgtatataca atatttctag
     7321 tttgttaaag agaattaaga aaataaatct cgaaaataat aaagggaaaa tcagtttttg
     7381 atatcaaaat tatacatgtc aacgataata caaaatataa tacaaactat aagatgttat
     7441 cagtatttat tatcatttag aataaatttt gtgtcgccct taattgtgag cggataacaa
     7501 ttacgagctt catgcacagt ggcgttgact caatattggc cattagccat attattcatt
     7561 ggttatatag cataaatcaa tattggctat tggccattgc atacgttgta tctatatcat
     7621 aatatgtaca tttatattgg ctcatgtcca atatgaccgc catgtt
//
