LOCUS       sequence-368853-        6496 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 6496)
  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 6496)
  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-SCN2B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     regulatory      2097..2106
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(2218..2237)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2250..2268)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2251..2268)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2478..2502)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2499..2702)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2528..2548)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2703..3082
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3249..3265
                     /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)."
     primer_bind     complement(3721..3738)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3721..3737)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3836..3853)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4007..4595)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4087..4106)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4681..4700)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4706..4725)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4767..4858
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4859..4927,4928..5719)
                     /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     complement(5077..5096)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5772..5893
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5783..5802)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5837..5856
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6023..6496,1..126))
                     /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(6106..6126)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc agccacggtg ctgtccctct caggaggttc
     1741 ttccatcagg acctgcagat ggatctttcc gtggcctctg tgccggtctg gagggttcat
     1801 gatgtagcag ttgtagatgc cctcgtcctc aggctgcacg tttctcagca tcacggacac
     1861 gtcgtactta gaggggttgc cgctgaattc cactctgtcc tggaatcttt ccagcttcag
     1921 gttgatgatc ttcatccgga actgcaggaa catttcctcg ctgcagttgt tgcactcttg
     1981 gtaggtccag ttcagggaga actgcttgtg gttcacggtg tagcagctat tgaaggtgca
     2041 gggcagtctg gcatcagagc cattcagcac gttcagtgtg gcaggcactg taacttccat
     2101 ggcggccgca gcgaacacca ggcagaagat gtagctcagg gcaatgatgg tcttcatggt
     2161 ggcgctagca ggcgcgtttt cgatagaacc gaggtgcagt tggacctggg agtggacacc
     2221 tgtggagaga aaggcaaagt ggatgttatt ctatagtgtc acctaaatcg tatgtgtatg
     2281 atacataagg ttatgtatta attgtagccg cgttctaacg aagccaaggg ggtgggccta
     2341 tagactctat aggcggtact tacgtcactc ttggcacggg gaatccgcgt tccaatgcac
     2401 cgttcccggc cgcggaggct ggatcggtcc cggtgtcttc tatggaggtc aaaacagcgt
     2461 ggatggcgtc tccaggcgat ctgacggttc actaaacgag ctctgcttat atagacctcc
     2521 caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg acattttgga
     2581 aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg gtggagactt
     2641 ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca aaaccgcatc
     2701 accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa gtcccataag
     2761 gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc aatagggggc
     2821 gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg taaatactcc
     2881 acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata cgtcattatt
     2941 gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac cgtaagttat
     3001 gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga ttactattaa
     3061 taactagtca ataatcaatg ccaacatggc ggtcatattg gacatgagcc aatataaatg
     3121 tacatattat gatatagata caacgtatgc aatggccaat agccaatatt gatttatgct
     3181 atataaccaa tgaataatat ggctaatggc caatattgag tcaacgccac tgtgcatgaa
     3241 gctcgtaatt gttatccgct cacaattaag ggcgacacaa aatttattct aaatgataat
     3301 aaatactgat aacatcttat agtttgtatt atattttgta ttatcgttga catgtataat
     3361 tttgatatca aaaactgatt ttccctttat tattttcgag atttattttc ttaattctct
     3421 ttaacaaact agaaatattg tatatacaaa aaatcataaa taatagatga atagtttaat
     3481 tataggtgtt catcaatcga aaaagcaacg tatcttattt aaagtgcgtt gcttttttct
     3541 catttataag gttaaataat tctcatatat caagcaaagt gacaggcgcc cttaaatatt
     3601 ctgacaaatg ctctttccct aaactccccc cataaaaaaa cccgccgaag cgggttttta
     3661 cgttatttgc ggattaacga ttactcgtta tcagaaccgc ccagggggcc cgagcttaag
     3721 actggccgtc gttttacaac acagaaagag tttgtagaaa cgcaaaaagg ccatccgtca
     3781 ggggccttct gcttagtttg atgcctggca gttccctact ctcgccttcc gcttcctcgc
     3841 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     3901 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     3961 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4021 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4081 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4141 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4201 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4261 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4321 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4381 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtgggctaac tacggctaca
     4441 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     4501 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     4561 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     4621 ggtctgacgc tcagtggaac gacgcgcgcg taactcacgt taagggattt tggtcatgag
     4681 caaggggtgt tatgagccat attcaggtat aaatgggctc gcgataatgt tcagaattgg
     4741 ttaattggtt gtaacactga cccctatttg tttatttttc taaatacatt caaatatgta
     4801 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagaatat
     4861 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4921 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4981 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     5041 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     5101 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5161 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5221 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5281 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5341 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5401 tgtagcgatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5461 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5521 ggcccttccg gctggctggt ttattgctga taaatccgga gccggtgagc gtggttctcg
     5581 cggtatcatc gcagcgctgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5641 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5701 actgattaag cattggtaat gaccatgatt acgccaagct ctagctagag gtcgacggta
     5761 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     5821 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     5881 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     5941 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     6001 aatcgaaatc tcgtagcacg tgtcaggcac cgggcttgcg ggtcatgcac caggtgcgcg
     6061 gtccttcggg cacctcgacg tcggcggtga cggtgaagcc gagccgctcg tagaagggga
     6121 ggttgcgggg cgcggatgtc tccaggaagg cgggcacccc ggcgcgctcg gccgcctcca
     6181 ctccggggag cacgacggcg ctgcccagac ccttgccctg gtggtcgggc gagacgccga
     6241 cggtggccag gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct
     6301 gttgctgcgc ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga
     6361 acaccgcccc cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt
     6421 ccgcgaccca caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct
     6481 cggtgacccg ctcgat
//
