LOCUS       sequence-368609-        6532 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 6532)
  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 6532)
  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-SCN1B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     181..201
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    319..430
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     419..436
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(576..593)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(594..638)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1881..1900)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1913..1931)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1914..1931)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2141..2165)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2162..2365)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2191..2211)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2366..2745
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2912..2928
                     /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(3384..3401)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3384..3400)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3499..3516)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3670..4258)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3750..3769)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4344..4363)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4369..4388)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4430..4521
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4522..4590,4591..5382)
                     /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(4740..4759)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5435..5556
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5446..5465)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5500..5519
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5686..6285)
                     /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(5769..5789)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6266..6285
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6304..6351)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6399..6532,1..196))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6413..6532,1..16)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6467..6486)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac taattgagat
       61 gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac ctggttgctg
      121 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
      181 accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt aaatcagctc
      241 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      301 gatagggttg agtgttgtga tgcaatttcc tcattttatt aggaaaggac agtgggagtg
      361 gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg ctggcaacta
      421 gaaggcacag tcgaggctga ttttctctat ctatttattt tactactcaa gactatcttg
      481 caaagtagaa gaaccctgtc tgcagtatga cgaccgtgta tacccgattg gcttaccggg
      541 taacgcgttg tgccagtgac tacaggccgg ccttagtggt gatggtggtg gtgttcatgc
      601 cactcaatct tctgagcctc gaaaatatca ttaagaccgc tgccgccggg gctcaggctc
      661 agggacttct gggtgtagtg gttgtgcagg gcctcgtgca tcacgctgca gctgaacacg
      721 ttgccctgct gccacctgga cttgtccacg gtcagcttgc tgtacaggaa gaagctgccg
      781 tcgctgtcca gcactggggg ggtggtcttg tagttgttct cgggctggcc gttgctctcc
      841 cactccacgg cgatatcgct ggggtagaag cccttcacca gacaggtcag ggacacctgg
      901 ttcttggtca tctcctcccg gctggggggc agggtgtaca cctggggctc ccgtggctgg
      961 cccttggcct tgctgattgt cttttcgatt ggggcagcca gggccttgtt ggagactttg
     1021 cacttgtatt ctttgccgtt cagccagtcc tggtgcagca cggtcagcac ggacaccacc
     1081 ctgtaggtgc tgttgtactg ctcctctctg ggcttggtct tggcgttgtg cacctccacg
     1141 ccgtccacgt accagttgaa cttcacctct gggtcctcgt gggacacggc caccaccacg
     1201 caggtcacct cgggggtcct gctgatcatc agggtgtcct tgggcttggg ggggaacagg
     1261 aacacggaag gccctcccag cagttctgga gctgggcagg gggggcaggt gtgggtcttg
     1321 tcgccgcccg gcgcgccttc ggacacgatg ctggccatgt ctctgttggc cttgtccacc
     1381 acctcgatgt ggattttctt gaccacgctg gtgttgtgct cgtagttctc gaagaacagc
     1441 agtctgtaca cgtggcactc gtagtcgccg ctgtggttgt aggtcacgtt ggtgatgaag
     1501 atgctcaggt cctgcagatc cttggtgcct ctgctgccat tccacacgac tctgccctca
     1561 aatctctcgt cctcttccag ctgcagcacc tcgttctcgt atctcaggat cttcacgaat
     1621 tcctcggtgc ccttctgccg gaaggtccac tcggtgaatg tctcggcatt tgtctcgctc
     1681 cgccgcttgc agctgatgca caggatcttg aaggtcatgc cgtacacggc ctctgtctca
     1741 gaatccactt ccacacatcc accggcggcc gcagcgaaca ccaggcagaa gatgtagctc
     1801 agggcaatga tggtcttcat ggtggcgcta gcaggcgcgt tttcgataga accgaggtgc
     1861 agttggacct gggagtggac acctgtggag agaaaggcaa agtggatgtt attctatagt
     1921 gtcacctaaa tcgtatgtgt atgatacata aggttatgta ttaattgtag ccgcgttcta
     1981 acgaagccaa gggggtgggc ctatagactc tataggcggt acttacgtca ctcttggcac
     2041 ggggaatccg cgttccaatg caccgttccc ggccgcggag gctggatcgg tcccggtgtc
     2101 ttctatggag gtcaaaacag cgtggatggc gtctccaggc gatctgacgg ttcactaaac
     2161 gagctctgct tatatagacc tcccaccgta cacgcctacc gcccatttgc gtcaatgggg
     2221 cggagttgtt acgacatttt ggaaagtccc gttgattttg gtgccaaaac aaactcccat
     2281 tgacgtcaat ggggtggaga cttggaaatc cccgtgagtc aaaccgctat ccacgcccat
     2341 tgatgtactg ccaaaaccgc atcaccatgg taatagcgat gactaatacg tagatgtact
     2401 gccaagtagg aaagtcccat aaggtcatgt actgggcata atgccaggcg ggccatttac
     2461 cgtcattgac gtcaataggg ggcgtacttg gcatatgata cacttgatgt actgccaagt
     2521 gggcagttta ccgtaaatac tccacccatt gacgtcaatg gaaagtccct attggcgtta
     2581 ctatgggaac atacgtcatt attgacgtca atgggcgggg gtcgttgggc ggtcagccag
     2641 gcgggccatt taccgtaagt tatgtaacgc ggaactccat atatgggcta tgaactaatg
     2701 accccgtaat tgattactat taataactag tcaataatca atgccaacat ggcggtcata
     2761 ttggacatga gccaatataa atgtacatat tatgatatag atacaacgta tgcaatggcc
     2821 aatagccaat attgatttat gctatataac caatgaataa tatggctaat ggccaatatt
     2881 gagtcaacgc cactgtgcat gaagctcgta attgttatcc gctcacaatt aagggcgaca
     2941 caaaatttat tctaaatgat aataaatact gataacatct tatagtttgt attatatttt
     3001 gtattatcgt tgacatgtat aattttgata tcaaaaactg attttccctt tattattttc
     3061 gagatttatt ttcttaattc tctttaacaa actagaaata ttgtatatac aaaaaatcat
     3121 aaataataga tgaatagttt aattataggt gttcatcaat cgaaaaagca acgtatctta
     3181 tttaaagtgc gttgcttttt tctcatttat aaggttaaat aattctcata tatcaagcaa
     3241 agtgacaggc gcccttaaat attctgacaa atgctctttc cctaaactcc ccccataaaa
     3301 aaacccgccg aagcgggttt ttacgttatt tgcggattaa cgattactcg ttatcagaac
     3361 cgcccagggg gcccgagctt aagactggcc gtcgttttac aacacagaaa gagtttgtag
     3421 aaacgcaaaa aggccatccg tcaggggcct tctgcttagt ttgatgcctg gcagttccct
     3481 actctcgcct tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg
     3541 agcggtatca gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc
     3601 aggaaagaac atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt
     3661 gctggcgttt ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag
     3721 tcagaggtgg cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc
     3781 cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc
     3841 ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt
     3901 cgttcgctcc aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt
     3961 atccggtaac tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc
     4021 agccactggt aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa
     4081 gtggtgggct aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa
     4141 gccagttacc ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg
     4201 tagcggtggt ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga
     4261 agatcctttg atcttttcta cggggtctga cgctcagtgg aacgacgcgc gcgtaactca
     4321 cgttaaggga ttttggtcat gagcaagggg tgttatgagc catattcagg tataaatggg
     4381 ctcgcgataa tgttcagaat tggttaattg gttgtaacac tgacccctat ttgtttattt
     4441 ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata aatgcttcaa
     4501 taatattgaa aaaggaagaa tatgagtatt caacatttcc gtgtcgccct tattcccttt
     4561 tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa agtaaaagat
     4621 gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa cagcggtaag
     4681 atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt taaagttctg
     4741 ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg tcgccgcata
     4801 cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca tcttacggat
     4861 ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa cactgcggcc
     4921 aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt gcacaacatg
     4981 ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc cataccaaac
     5041 gacgagcgtg acaccacgat gcctgtagcg atggcaacaa cgttgcgcaa actattaact
     5101 ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga ggcggataaa
     5161 gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc tgataaatcc
     5221 ggagccggtg agcgtggttc tcgcggtatc atcgcagcgc tggggccaga tggtaagccc
     5281 tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga acgaaataga
     5341 cagatcgctg agataggtgc ctcactgatt aagcattggt aatgaccatg attacgccaa
     5401 gctctagcta gaggtcgacg gtatacagac atgataagat acattgatga gtttggacaa
     5461 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     5521 ttatttgtaa ccattataag ctgcaataaa caagttgggg tgggcgaaga actccagcat
     5581 gagatccccg cgctggagga tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa
     5641 cctttcatag aaggcggcgg tggaatcgaa atctcgtagc acgtgtcagg caccgggctt
     5701 gcgggtcatg caccaggtgc gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa
     5761 gccgagccgc tcgtagaagg ggaggttgcg gggcgcggat gtctccagga aggcgggcac
     5821 cccggcgcgc tcggccgcct ccactccggg gagcacgacg gcgctgccca gacccttgcc
     5881 ctggtggtcg ggcgagacgc cgacggtggc caggaaccac gcgggctcct tgggccggtg
     5941 cggcgccagg aggccttcca tctgttgctg cgcggccagc cgggaaccgc tcaactcggc
     6001 catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg acgctctccg gcgtggtcca
     6061 gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg
     6121 cgtgaggaag agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt
     6181 gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac
     6241 gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt agttcctcac
     6301 cttgtcgtat tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga
     6361 tcagatccga aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca
     6421 aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca
     6481 taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg cg
//
