LOCUS       sequence-368854-        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-CD160-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6496
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     191..211
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    329..440
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     429..446
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(586..603)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(604..648)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1855..1874)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1887..1905)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1888..1905)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2115..2139)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2136..2339)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2165..2185)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2340..2719
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2886..2902
                     /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(3358..3375)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3358..3374)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3473..3490)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3644..4232)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3724..3743)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4318..4337)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4343..4362)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4404..4495
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4496..4564,4565..5356)
                     /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(4714..4733)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5409..5530
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5420..5439)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5474..5493
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5660..6259)
                     /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(5743..5763)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6240..6259
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6278..6325)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6373..6496,1..206))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6387..6496,1..26)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gaactgggcg gagttagggg cgggatgggc ggagttaggg gcgggactat ggttgctgac
       61 taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga ctttccacac
      121 ctggttgctg actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg
      181 gactttccac accctaactg acacacattc cacagaatta attcgcgtta aatttttgtt
      241 aaatcagctc attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag
      301 aatagaccga gatagggttg agtgttgtga tgcaatttcc tcattttatt aggaaaggac
      361 agtgggagtg gcaccttcca gggtcaagga aggcacgggg gaggggcaaa caacagatgg
      421 ctggcaacta gaaggcacag tcgaggctga ttttctctat ctatttattt tactactcaa
      481 gactatcttg caaagtagaa gaaccctgtc tgcagtatga cgaccgtgta tacccgattg
      541 gcttaccggg taacgcgttg tgccagtgac tacaggccgg ccttagtggt gatggtggtg
      601 gtgttcatgc cactcaatct tctgagcctc gaaaatatca ttaagaccgc tgccgccggg
      661 gctcaggctc agggacttct gggtgtagtg gttgtgcagg gcctcgtgca tcacgctgca
      721 gctgaacacg ttgccctgct gccacctgga cttgtccacg gtcagcttgc tgtacaggaa
      781 gaagctgccg tcgctgtcca gcactggggg ggtggtcttg tagttgttct cgggctggcc
      841 gttgctctcc cactccacgg cgatatcgct ggggtagaag cccttcacca gacaggtcag
      901 ggacacctgg ttcttggtca tctcctcccg gctggggggc agggtgtaca cctggggctc
      961 ccgtggctgg cccttggcct tgctgattgt cttttcgatt ggggcagcca gggccttgtt
     1021 ggagactttg cacttgtatt ctttgccgtt cagccagtcc tggtgcagca cggtcagcac
     1081 ggacaccacc ctgtaggtgc tgttgtactg ctcctctctg ggcttggtct tggcgttgtg
     1141 cacctccacg ccgtccacgt accagttgaa cttcacctct gggtcctcgt gggacacggc
     1201 caccaccacg caggtcacct cgggggtcct gctgatcatc agggtgtcct tgggcttggg
     1261 ggggaacagg aacacggaag gccctcccag cagttctgga gctgggcagg gggggcaggt
     1321 gtgggtcttg tcgccgcccg gcgcgccacc ttcgttgtgg ctgaattcca ggtgctgccg
     1381 ctgtttcagt ccggtcacgg tgtagttgcc tgtctcggtg aacaggatgc tgaagaagtg
     1441 gccctgcagt ctgatgccgg acttctgaga tctggcgcaa cactggtagg tgccgctgtg
     1501 cagaggtgtc acttggctga tggtgaacat cagctggcta gagatctcgc ccacgccgtc
     1561 gatgccagga tctctcttca gtctcagctg cttcaggctt gtctcagggc tgcagtcgcc
     1621 gcttctatcc ttgcacagga acaccacgaa gccctcggcc tcttctttct tgtgccacac
     1681 ggtgcagatc agattcagtc tggtgccctc ttgagaggcg ctgcttgtga tattgatggc
     1741 ggccgcagcg aacaccaggc agaagatgta gctcagggca atgatggtct tcatggtggc
     1801 gctagcaggc gcgttttcga tagaaccgag gtgcagttgg acctgggagt ggacacctgt
     1861 ggagagaaag gcaaagtgga tgttattcta tagtgtcacc taaatcgtat gtgtatgata
     1921 cataaggtta tgtattaatt gtagccgcgt tctaacgaag ccaagggggt gggcctatag
     1981 actctatagg cggtacttac gtcactcttg gcacggggaa tccgcgttcc aatgcaccgt
     2041 tcccggccgc ggaggctgga tcggtcccgg tgtcttctat ggaggtcaaa acagcgtgga
     2101 tggcgtctcc aggcgatctg acggttcact aaacgagctc tgcttatata gacctcccac
     2161 cgtacacgcc taccgcccat ttgcgtcaat ggggcggagt tgttacgaca ttttggaaag
     2221 tcccgttgat tttggtgcca aaacaaactc ccattgacgt caatggggtg gagacttgga
     2281 aatccccgtg agtcaaaccg ctatccacgc ccattgatgt actgccaaaa ccgcatcacc
     2341 atggtaatag cgatgactaa tacgtagatg tactgccaag taggaaagtc ccataaggtc
     2401 atgtactggg cataatgcca ggcgggccat ttaccgtcat tgacgtcaat agggggcgta
     2461 cttggcatat gatacacttg atgtactgcc aagtgggcag tttaccgtaa atactccacc
     2521 cattgacgtc aatggaaagt ccctattggc gttactatgg gaacatacgt cattattgac
     2581 gtcaatgggc gggggtcgtt gggcggtcag ccaggcgggc catttaccgt aagttatgta
     2641 acgcggaact ccatatatgg gctatgaact aatgaccccg taattgatta ctattaataa
     2701 ctagtcaata atcaatgcca acatggcggt catattggac atgagccaat ataaatgtac
     2761 atattatgat atagatacaa cgtatgcaat ggccaatagc caatattgat ttatgctata
     2821 taaccaatga ataatatggc taatggccaa tattgagtca acgccactgt gcatgaagct
     2881 cgtaattgtt atccgctcac aattaagggc gacacaaaat ttattctaaa tgataataaa
     2941 tactgataac atcttatagt ttgtattata ttttgtatta tcgttgacat gtataatttt
     3001 gatatcaaaa actgattttc cctttattat tttcgagatt tattttctta attctcttta
     3061 acaaactaga aatattgtat atacaaaaaa tcataaataa tagatgaata gtttaattat
     3121 aggtgttcat caatcgaaaa agcaacgtat cttatttaaa gtgcgttgct tttttctcat
     3181 ttataaggtt aaataattct catatatcaa gcaaagtgac aggcgccctt aaatattctg
     3241 acaaatgctc tttccctaaa ctccccccat aaaaaaaccc gccgaagcgg gtttttacgt
     3301 tatttgcgga ttaacgatta ctcgttatca gaaccgccca gggggcccga gcttaagact
     3361 ggccgtcgtt ttacaacaca gaaagagttt gtagaaacgc aaaaaggcca tccgtcaggg
     3421 gccttctgct tagtttgatg cctggcagtt ccctactctc gccttccgct tcctcgctca
     3481 ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac tcaaaggcgg
     3541 taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga gcaaaaggcc
     3601 agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat aggctccgcc
     3661 cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac ccgacaggac
     3721 tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct gttccgaccc
     3781 tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg ctttctcata
     3841 gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg ggctgtgtgc
     3901 acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt cttgagtcca
     3961 acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg attagcagag
     4021 cgaggtatgt aggcggtgct acagagttct tgaagtggtg ggctaactac ggctacacta
     4081 gaagaacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga aaaagagttg
     4141 gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt gtttgcaagc
     4201 agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt tctacggggt
     4261 ctgacgctca gtggaacgac gcgcgcgtaa ctcacgttaa gggattttgg tcatgagcaa
     4321 ggggtgttat gagccatatt caggtataaa tgggctcgcg ataatgttca gaattggtta
     4381 attggttgta acactgaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     4441 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agaatatgag
     4501 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     4561 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     4621 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     4681 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     4741 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     4801 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     4861 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     4921 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     4981 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     5041 agcgatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     5101 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     5161 ccttccggct ggctggttta ttgctgataa atccggagcc ggtgagcgtg gttctcgcgg
     5221 tatcatcgca gcgctggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     5281 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     5341 gattaagcat tggtaatgac catgattacg ccaagctcta gctagaggtc gacggtatac
     5401 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     5461 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     5521 taaacaagtt ggggtgggcg aagaactcca gcatgagatc cccgcgctgg aggatcatcc
     5581 agccggcgtc ccggaaaacg attccgaagc ccaacctttc atagaaggcg gcggtggaat
     5641 cgaaatctcg tagcacgtgt caggcaccgg gcttgcgggt catgcaccag gtgcgcggtc
     5701 cttcgggcac ctcgacgtcg gcggtgacgg tgaagccgag ccgctcgtag aaggggaggt
     5761 tgcggggcgc ggatgtctcc aggaaggcgg gcaccccggc gcgctcggcc gcctccactc
     5821 cggggagcac gacggcgctg cccagaccct tgccctggtg gtcgggcgag acgccgacgg
     5881 tggccaggaa ccacgcgggc tccttgggcc ggtgcggcgc caggaggcct tccatctgtt
     5941 gctgcgcggc cagccgggaa ccgctcaact cggccatgcg cgggccgatc tcggcgaaca
     6001 ccgcccccgc ttcgacgctc tccggcgtgg tccagaccgc caccgcggcg ccgtcgtccg
     6061 cgacccacac cttgccgatg tcgagcccga cgcgcgtgag gaagagttct tgcagctcgg
     6121 tgacccgctc gatgtggcgg tccggatcga cggtgtggcg cgtggcgggg tagtcggcga
     6181 acgcggcggc gagggtgcgt acggccctgg ggacgtcgtc gcgggtggcg aggcgcaccg
     6241 tgggcttgta ctcggtcatg gtttagttcc tcaccttgtc gtattatact atgccgatat
     6301 actatgccga tgattaattg tcaacacgtg ctgatcagat ccgaaaatgg atatacaagc
     6361 tcccgggagc tttttgcaaa agcctaggcc tccaaaaaag cctcctcact acttctggaa
     6421 tagctcagag gcagaggcgg cctcggcctc tgcataaata aaaaaaatta gtcagccatg
     6481 gggcggagaa tgggcg
//
