LOCUS       sequence-303129-        8971 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein partial 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 8971)
  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 8971)
  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-PTPRQ(Trunc3)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8971
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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)."
     enhancer        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             6313..6357
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6358..6375
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6515..6532)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6521..6632
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6750..6770)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6755..7084
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6935..7070
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6997..7016
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7132..7179
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7198..7797
                     /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(7198..7217)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7694..7714
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7927..8048
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7964..7983)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8018..8037
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8101..8892,8893..8961))
                     /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     8724..8743
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8962..8971,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgcccc tccaaaggat
     2761 gtgtactacg ccaatctgag cagcagctcc atcatcctgt tctggacccc tccaagcaag
     2821 cccaacggca tcatccagta ctacagcgtg tactaccgga acaccagcgg caccttcatg
     2881 cagaacttca ccctgcacga agtgaccaac gacttcgaca acatgaccgt gtccaccatc
     2941 atcgacaagc tgaccatctt cagctactac accttctggc tgaccgccag cacaagcgtc
     3001 ggcaatggca acaagagcag cgacatcatc gaggtgtaca cagaccagga catccctgag
     3061 ggcttcgtgg gcaatctgac ctacgagagc atcagcagca ccgccatcaa tgtgtcttgg
     3121 gtgccaccag ctcagcccaa tggcctggtg ttttactacg tgtccctgat cctgcagcag
     3181 acccctagac atgttcggcc tccactggtc acatacgagc ggagcatcta cttcgataac
     3241 ctcgagaagt acaccgacta catcctgaag atcaccccta gcaccgagaa gggcttcagc
     3301 gatacctaca cagcccagct gtacatcaag accgaagagg acgtgcccga gacaagcccc
     3361 atcatcaaca ccttcaagaa cctgtccagc accagcgtgc tgctgtcttg ggaccctcct
     3421 gtgaagccta acggcgccat catctcctac gatctgacac tgcagggccc caacgagaac
     3481 tacagcttca tcaccagcga caactacatc attctggaag aactgagccc cttcacgctc
     3541 tacagctttt ttgccgccgc tcggaccaga aaaggcctgg gacctagcag catcctgttt
     3601 ttctacaccg acgagagcgt gcccctggct cctcctcaga atctgaccct gatcaactgc
     3661 acctccgact tcgtgtggct gaagtggtcc ccttctcctc tgcctggcgg aatcgtgaag
     3721 gtgtacagct ttaagatcca cgagcacgag acagacacca tctactacaa gaacatcagc
     3781 ggctttaaga ccgaggccaa gctcgtggga ctcgagcctg tgtctaccta cagcatcaga
     3841 gtgtccgcct tcaccaaagt tggcaacggc aaccagttca gcaacgtggt caagttcacc
     3901 acacaagagt ccgtgcctga cgtggtgcag aacatgcagt gtatggccac aagctggcag
     3961 agcgtgctgg ttaagtggga cccacctaag aaggccaacg gaatcattac ccagtacatg
     4021 gtcaccgtgg aacggaacag caccaaggtg tccccacagg accacatgta cacattcatc
     4081 aagctgctgg ccaacaccag ctacgtgttc aaagtgcggg cctctacatc tgccggcgag
     4141 ggcgacgaaa gcacatgtca tgtgtccaca ctgcctgaga cagtgcccag cgtgcccacc
     4201 aatatcgcct tttccgatgt gcagagcacc tccgccacac tgacctggat cagacctgat
     4261 accatcctgg gctacttcca gaactacaag atcacaaccc agctgcgggc ccagaaatgc
     4321 aaagagtggg agagcgagga atgcgtcgag taccagaaga tccagtacct gtacgaggcc
     4381 cacctgaccg aggaaaccgt gtacggcctg aagaaattcc ggtggtacag attccaggtg
     4441 gccgcctcta caaacgccgg ctacggaaat gccagcaact ggatcagcac aaagaccctg
     4501 cctggacctc ctgacggccc tcctgaaaat gtgcatgtgg tggccacatc tcccttcagc
     4561 atctccatct cttggagcga gcctgccgtg atcacaggcc ctacctgtta cctgatcgac
     4621 gtgaagtccg tggacaacga cgagttcaac atctccttca tcaagagcaa cgaggaaaac
     4681 aagaccatcg agatcaagga cctggaaatc ttcacccggt acagcgtggt catcaccgcc
     4741 tttacaggca atatcagcgc cgcctacgtg gaaggcaagt cctctgccga gatgatcgtg
     4801 accacactgg aaagcgcccc taaggaccct ccaaacaata tgaccttcca gaaaatcccc
     4861 gacgaagtca ccaagttcca gctgaccttc ctgcctccta gccagcctaa tggcaatatc
     4921 caggtgtacc aggctctggt gtacagagag gatgatccca cagccgtgca gatccacaac
     4981 ctgtctatca tccagaaaac gaacaccttc gtgatcgcca tgctggaagg actgaaaggc
     5041 gggcacacct acaatatctc cgtgtacgcc gtgaatagcg ctggcgcagg acctaaggtg
     5101 cccatgagaa tcaccatgga catcaaggcc cctgccagac ctaagaccaa gcctacacct
     5161 atctacgacg ccaccggcaa gctgctcgtg accagcacca caatcaccat cagaatgccc
     5221 atctgctact actccgacga tcacggcccc atcaagaacg tgcaggtcct ggttacagaa
     5281 acaggcgccc agcacgacgg caacgtgaca aagtggtacg acgcctactt caacaaggcc
     5341 agaccttact tcaccaacga gggcttcccc aatcctcctt gtaccgaggg caagaccaag
     5401 ttcagcggca acgaagagat ctatatcatc ggagccgaca acgcctgcat gatccccggc
     5461 aatgaggaca agatctgcaa tggccctctg aagcccaaga agcagtatct gttcaagttc
     5521 cgggccacca acatcatggg ccagttcacc gacagcgact acagcgaccc tgtgaaaaca
     5581 ctcggcgagg gactgagcga gagaaccgtt gaaggcgcgc cgggcggcga caagacccac
     5641 acctgccccc cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc
     5701 cccaagccca aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg
     5761 gccgtgtccc acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg
     5821 cacaacgcca agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc
     5881 gtgctgaccg tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc
     5941 aacaaggccc tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg
     6001 gagccccagg tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc
     6061 ctgacctgtc tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac
     6121 ggccagcccg agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc
     6181 ttcctgtaca gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc
     6241 tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc
     6301 cccggcggca gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac
     6361 caccaccatc accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc
     6421 caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc
     6481 ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag
     6541 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6601 gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt
     6661 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6721 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6781 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6841 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6901 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6961 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     7021 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     7081 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     7141 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     7201 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     7261 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     7321 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     7381 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     7441 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     7501 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     7561 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     7621 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     7681 acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     7741 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac
     7801 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7861 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7921 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7981 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     8041 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     8101 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     8161 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     8221 cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa
     8281 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8341 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8401 cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8461 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8521 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8581 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8641 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8701 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8761 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8821 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8881 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8941 acggaaatgt tgaatactca tattcttcct t
//
