LOCUS       sequence-368812-        6430 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 6430)
  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 6430)
  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-THY1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6430
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     164..184
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    302..413
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     402..419
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(559..576)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(577..621)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1762..1781)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1794..1812)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1795..1812)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2022..2046)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2043..2246)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2072..2092)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2247..2626
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2793..2809
                     /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(3265..3282)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3265..3281)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3380..3397)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3551..4139)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3631..3650)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4225..4244)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4250..4269)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4311..4402
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4403..4471,4472..5263)
                     /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(4621..4640)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5316..5437
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5327..5346)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5381..5400
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5567..6166)
                     /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(5650..5670)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6147..6166
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6185..6232)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6280..6430,1..179))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6294..6429
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6348..6367)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct ttgcatactt
       61 ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt gagatgcatg
      121 ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa ctgacacaca
      181 ttccacagaa ttaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg
      241 ccgaaatcgg caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg
      301 tgatgcaatt tcctcatttt attaggaaag gacagtggga gtggcacctt ccagggtcaa
      361 ggaaggcacg ggggaggggc aaacaacaga tggctggcaa ctagaaggca cagtcgaggc
      421 tgattttctc tatctattta ttttactact caagactatc ttgcaaagta gaagaaccct
      481 gtctgcagta tgacgaccgt gtatacccga ttggcttacc gggtaacgcg ttgtgccagt
      541 gactacaggc cggccttagt ggtgatggtg gtggtgttca tgccactcaa tcttctgagc
      601 ctcgaaaata tcattaagac cgctgccgcc ggggctcagg ctcagggact tctgggtgta
      661 gtggttgtgc agggcctcgt gcatcacgct gcagctgaac acgttgccct gctgccacct
      721 ggacttgtcc acggtcagct tgctgtacag gaagaagctg ccgtcgctgt ccagcactgg
      781 gggggtggtc ttgtagttgt tctcgggctg gccgttgctc tcccactcca cggcgatatc
      841 gctggggtag aagcccttca ccagacaggt cagggacacc tggttcttgg tcatctcctc
      901 ccggctgggg ggcagggtgt acacctgggg ctcccgtggc tggcccttgg ccttgctgat
      961 tgtcttttcg attggggcag ccagggcctt gttggagact ttgcacttgt attctttgcc
     1021 gttcagccag tcctggtgca gcacggtcag cacggacacc accctgtagg tgctgttgta
     1081 ctgctcctct ctgggcttgg tcttggcgtt gtgcacctcc acgccgtcca cgtaccagtt
     1141 gaacttcacc tctgggtcct cgtgggacac ggccaccacc acgcaggtca cctcgggggt
     1201 cctgctgatc atcagggtgt ccttgggctt gggggggaac aggaacacgg aaggccctcc
     1261 cagcagttct ggagctgggc agggggggca ggtgtgggtc ttgtcgccgc ccggcgcgcc
     1321 aagtttgtcc cgcagcacgg tcacgttctg gctgctgatt ggagggctgt gtccagagtg
     1381 gtgcagggcg catgtgtatg tgccctcatc cttggaggtg aaggcgctca ggtacagcac
     1441 cttcatgttg tacttgctgg tgaagttggt cctgcttctg tatgtgtgct ctggcacgcc
     1501 cacggtgccg aacagcacgt gtttctttgt ttctctggtc aggctgaact cgtactggat
     1561 ggggctgctg ctggtattct cgtgtctgca gtccagtctc aggctctggt ccaccaggca
     1621 agctgtcaga gatgtcactt tctgggcggc cgcagcgaac accaggcaga agatgtagct
     1681 cagggcaatg atggtcttca tggtggcgct agcaggcgcg ttttcgatag aaccgaggtg
     1741 cagttggacc tgggagtgga cacctgtgga gagaaaggca aagtggatgt tattctatag
     1801 tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct
     1861 aacgaagcca agggggtggg cctatagact ctataggcgg tacttacgtc actcttggca
     1921 cggggaatcc gcgttccaat gcaccgttcc cggccgcgga ggctggatcg gtcccggtgt
     1981 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa
     2041 cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2101 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2161 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2221 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2281 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     2341 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     2401 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     2461 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     2521 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     2581 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgccaaca tggcggtcat
     2641 attggacatg agccaatata aatgtacata ttatgatata gatacaacgt atgcaatggc
     2701 caatagccaa tattgattta tgctatataa ccaatgaata atatggctaa tggccaatat
     2761 tgagtcaacg ccactgtgca tgaagctcgt aattgttatc cgctcacaat taagggcgac
     2821 acaaaattta ttctaaatga taataaatac tgataacatc ttatagtttg tattatattt
     2881 tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct ttattatttt
     2941 cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca
     3001 taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt
     3061 atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca
     3121 aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa
     3181 aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa
     3241 ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta
     3301 gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc
     3361 tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3421 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3481 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3541 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3601 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3661 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3721 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3781 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3841 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3901 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3961 agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4021 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4081 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4141 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc
     4201 acgttaaggg attttggtca tgagcaaggg gtgttatgag ccatattcag gtataaatgg
     4261 gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt
     4321 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4381 ataatattga aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4441 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4501 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4561 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4621 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4681 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4741 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4801 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4861 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     4921 cgacgagcgt gacaccacga tgcctgtagc gatggcaaca acgttgcgca aactattaac
     4981 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5041 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5101 cggagccggt gagcgtggtt ctcgcggtat catcgcagcg ctggggccag atggtaagcc
     5161 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5221 acagatcgct gagataggtg cctcactgat taagcattgg taatgaccat gattacgcca
     5281 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     5341 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5401 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     5461 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     5521 acctttcata gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag gcaccgggct
     5581 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
     5641 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga tgtctccagg aaggcgggca
     5701 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
     5761 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
     5821 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
     5881 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
     5941 agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
     6001 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg
     6061 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga
     6121 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt tagttcctca
     6181 ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg
     6241 atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc
     6301 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc
     6361 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     6421 gggcgggatg
//
