LOCUS       sequence-368834-        6481 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 6481)
  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 6481)
  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-MPIG6B-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6481
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     114..133
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(152..199)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(247..576)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      261..396
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(315..334)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     561..581
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    699..810
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     799..816
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(956..973)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(974..1018)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2210..2229)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2242..2260)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2243..2260)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2470..2494)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2491..2694)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2520..2540)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2695..3074
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3241..3257
                     /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(3713..3730)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3713..3729)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3828..3845)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3999..4587)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4079..4098)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4673..4692)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4698..4717)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4759..4850
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4851..4919,4920..5711)
                     /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(5069..5088)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5764..5885
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5775..5794)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5829..5848
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6015..6481,1..133))
                     /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(6098..6118)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gctcgatgtg gcggtccgga tcgacggtgt ggcgcgtggc ggggtagtcg gcgaacgcgg
       61 cggcgagggt gcgtacggcc ctggggacgt cgtcgcgggt ggcgaggcgc accgtgggct
      121 tgtactcggt catggtttag ttcctcacct tgtcgtatta tactatgccg atatactatg
      181 ccgatgatta attgtcaaca cgtgctgatc agatccgaaa atggatatac aagctcccgg
      241 gagctttttg caaaagccta ggcctccaaa aaagcctcct cactacttct ggaatagctc
      301 agaggcagag gcggcctcgg cctctgcata aataaaaaaa attagtcagc catggggcgg
      361 agaatgggcg gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat
      421 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      481 ctttccacac ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg
      541 ggagcctggg gactttccac accctaactg acacacattc cacagaatta attcgcgtta
      601 aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat
      661 aaatcaaaag aatagaccga gatagggttg agtgttgtga tgcaatttcc tcattttatt
      721 aggaaaggac agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa
      781 caacagatgg ctggcaacta gaaggcacag tcgaggctga ttttctctat ctatttattt
      841 tactactcaa gactatcttg caaagtagaa gaaccctgtc tgcagtatga cgaccgtgta
      901 tacccgattg gcttaccggg taacgcgttg tgccagtgac tacaggccgg ccttagtggt
      961 gatggtggtg gtgttcatgc cactcaatct tctgagcctc gaaaatatca ttaagaccgc
     1021 tgccgccggg gctcaggctc agggacttct gggtgtagtg gttgtgcagg gcctcgtgca
     1081 tcacgctgca gctgaacacg ttgccctgct gccacctgga cttgtccacg gtcagcttgc
     1141 tgtacaggaa gaagctgccg tcgctgtcca gcactggggg ggtggtcttg tagttgttct
     1201 cgggctggcc gttgctctcc cactccacgg cgatatcgct ggggtagaag cccttcacca
     1261 gacaggtcag ggacacctgg ttcttggtca tctcctcccg gctggggggc agggtgtaca
     1321 cctggggctc ccgtggctgg cccttggcct tgctgattgt cttttcgatt ggggcagcca
     1381 gggccttgtt ggagactttg cacttgtatt ctttgccgtt cagccagtcc tggtgcagca
     1441 cggtcagcac ggacaccacc ctgtaggtgc tgttgtactg ctcctctctg ggcttggtct
     1501 tggcgttgtg cacctccacg ccgtccacgt accagttgaa cttcacctct gggtcctcgt
     1561 gggacacggc caccaccacg caggtcacct cgggggtcct gctgatcatc agggtgtcct
     1621 tgggcttggg ggggaacagg aacacggaag gccctcccag cagttctgga gctgggcagg
     1681 gggggcaggt gtgggtcttg tcgccgcccg gcgcgccttg agggtaaacg ctgccgtgtg
     1741 taggtccagg ggccttgcaa tatgttctgt cgcccagcac gtgcagcact gttctgctct
     1801 cgtcctcgtg cctgcctttg cagaaaaatg tgccagagtc gccagcagac agcagcagtt
     1861 ccagccgtct gatgccgcta tccagagatc tcagtctgcc cacgaagggc tgcagtggag
     1921 gaactgtagg tgttccagag ctagaggccc acaggatagg ccgtctgcct ttagaaaggc
     1981 ccttgcaggc aggaaagctt ggggcccaaa cccatctgat ggggtgagac actccgccac
     2041 agctcagatt cactctgtcg ccgggtctac catcaagaga agcaccagga ttggcggccg
     2101 cagcgaacac caggcagaag atgtagctca gggcaatgat ggtcttcatg gtggcgctag
     2161 caggcgcgtt ttcgatagaa ccgaggtgca gttggacctg ggagtggaca cctgtggaga
     2221 gaaaggcaaa gtggatgtta ttctatagtg tcacctaaat cgtatgtgta tgatacataa
     2281 ggttatgtat taattgtagc cgcgttctaa cgaagccaag ggggtgggcc tatagactct
     2341 ataggcggta cttacgtcac tcttggcacg gggaatccgc gttccaatgc accgttcccg
     2401 gccgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc gtggatggcg
     2461 tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct cccaccgtac
     2521 acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg gaaagtcccg
     2581 ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac ttggaaatcc
     2641 ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca tcaccatggt
     2701 aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata aggtcatgta
     2761 ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg gcgtacttgg
     2821 catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact ccacccattg
     2881 acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa
     2941 tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt atgtaacgcg
     3001 gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt aataactagt
     3061 caataatcaa tgccaacatg gcggtcatat tggacatgag ccaatataaa tgtacatatt
     3121 atgatataga tacaacgtat gcaatggcca atagccaata ttgatttatg ctatataacc
     3181 aatgaataat atggctaatg gccaatattg agtcaacgcc actgtgcatg aagctcgtaa
     3241 ttgttatccg ctcacaatta agggcgacac aaaatttatt ctaaatgata ataaatactg
     3301 ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata attttgatat
     3361 caaaaactga ttttcccttt attattttcg agatttattt tcttaattct ctttaacaaa
     3421 ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta attataggtg
     3481 ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt ctcatttata
     3541 aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata ttctgacaaa
     3601 tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt tacgttattt
     3661 gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta agactggccg
     3721 tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt caggggcctt
     3781 ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc gctcactgac
     3841 tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata
     3901 cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa
     3961 aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct
     4021 gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa
     4081 agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg
     4141 cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca
     4201 cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa
     4261 ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg
     4321 gtaagacacg acttatcgcc actggcagca gccactggta acaggattag cagagcgagg
     4381 tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta cactagaaga
     4441 acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc
     4501 tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag
     4561 attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac
     4621 gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg agcaaggggt
     4681 gttatgagcc atattcaggt ataaatgggc tcgcgataat gttcagaatt ggttaattgg
     4741 ttgtaacact gacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca
     4801 tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat atgagtattc
     4861 aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc
     4921 acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt
     4981 acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt
     5041 ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg
     5101 ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact
     5161 caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg
     5221 ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga
     5281 aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg
     5341 aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcga
     5401 tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac
     5461 aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc
     5521 cggctggctg gtttattgct gataaatccg gagccggtga gcgtggttct cgcggtatca
     5581 tcgcagcgct ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga
     5641 gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta
     5701 agcattggta atgaccatga ttacgccaag ctctagctag aggtcgacgg tatacagaca
     5761 tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga aaaaaatgct
     5821 ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc tgcaataaac
     5881 aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat catccagccg
     5941 gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt ggaatcgaaa
     6001 tctcgtagca cgtgtcaggc accgggcttg cgggtcatgc accaggtgcg cggtccttcg
     6061 ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg gaggttgcgg
     6121 ggcgcggatg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc cactccgggg
     6181 agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc gacggtggcc
     6241 aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat ctgttgctgc
     6301 gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc gaacaccgcc
     6361 cccgcttcga cgctctccgg cgtggtccag accgccaccg cggcgccgtc gtccgcgacc
     6421 cacaccttgc cgatgtcgag cccgacgcgc gtgaggaaga gttcttgcag ctcggtgacc
     6481 c
//
