LOCUS       sequence-303131-        8977 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 8977)
  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 8977)
  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(Trunc2)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8977
                     /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             6319..6363
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6364..6381
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6521..6538)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6527..6638
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6756..6776)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6761..7090
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6941..7076
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7003..7022
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7138..7185
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7204..7803
                     /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(7204..7223)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7700..7720
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7933..8054
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7970..7989)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8024..8043
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8107..8898,8899..8967))
                     /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     8730..8749
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8968..8977,1..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 cggccgccga aagataccaa
     2761 gtggacctgg tgcctgacag cggcttcgtg accatcagag atctcggcgg aggcgagtat
     2821 caggtggacg tgtcaaatgt ggtgcccggc acaagatacg acatcaccat cagcagcatc
     2881 agcaccacct acacaagccc cgtgaccaga atcgtgacca ccaacgtgac caagcctgga
     2941 cctcctgtgt ttctggccgg cgagagagtt ggatctgccg gcattctgct gagctggaac
     3001 acccctccta atcctaacgg ccggatcatc agctacatcg tgaagtacaa agaagtgtgc
     3061 ccctggatgc agaccgtgta cacacaagtg cggagcaagc ccgacagcct ggaagtgctg
     3121 ctgaccaacc tgaatcctgg cacaacctac gagatcaagg tggccgccga gaattctgcc
     3181 ggaatcggcg tgttcagcga ccccttcctg tttcaaaccg ccgaatctgc ccctggcaag
     3241 gtggtcaatc tgaccgtgga agcctacaac gccagcgccg tgaagctgat ctggtatctg
     3301 cccagacagc ccaacggcaa gatcaccagc ttcaagatca gcgtgaagca cgctcggagc
     3361 ggcatcgtgg tcaaggacgt gtccattaga gtggaagata tcctgaccgg caagctgccc
     3421 gagtgcaacg agaatagcga gagcttcctg tggtccaccg cctctccatc tccaacactg
     3481 ggcagagtga cccctccaag cagaaccaca cacagcagct ccacactgac ccagaacgag
     3541 atcagcagcg tgtggaaaga acccatcagc ttcgtggtca cccacctgag gccttacacc
     3601 acatacctgt tcgaggtgtc cgccgtgaca acagaggccg gctacatcga tagcaccatc
     3661 gtgcggacac cagagagcgt gccagaaggc cctcctcaga attgcgtgac aggcaacatc
     3721 accggcaaga gcttcagcat cctgtgggac cctccaacca tcgtgaccgg aaagttcagc
     3781 tacagagtgg aactgtacgg ccccagcggc agaatcctgg ataacagcac caaggacctg
     3841 aagttcgcct tcaccaatct gacacccttc actatgtacg atgtgtatat cgccgctgag
     3901 acaagcgccg gcacaggccc taagagcaac atcagcgtgt tcacccctcc agatgtgcct
     3961 ggcgccgtgt ttgatctgca gctggccgag gtggaatcta cccaagtccg gatcacctgg
     4021 aagaagccca ggcaacctaa cggcatcatc aaccagtatc gcgtgaaggt gctggtgccc
     4081 gagacaggca tcatcctgga aaacaccctg ctgaccggga acaacgagta catcaacgac
     4141 cccatggctc ccgagatcgt gaacatcgtg gaacccatgg tcggactgta cgagggctct
     4201 gccgagatgt ctagcgatct gcacagcctg gccacattca tctacaactc tcaccccgac
     4261 aagaacttcc ccgccagaaa tagagccgag gaccagacaa gccctgtggt caccaccaga
     4321 aaccagtaca tcaccgatat tgccgccgag cagctgagct acgtgatcag acgactggtg
     4381 cccttcaccg agcacatgat ctccgtgtcc gcctttacca tcatgggcga gggacctcca
     4441 acagtgctgt ctgtgcggac aagacaacag gtgcccagct ccatcaagat catcaattac
     4501 aagaacatca gctcctccag catcctcctg tactgggacc caccagagta ccctaatgga
     4561 aagatcaccc actacaccat ctacgccatg gaactggaca ccaaccgggc cttccagatc
     4621 acaacaatcg acaacagctt cctgatcacc ggcctgaaga agtacaccaa gtacaagatg
     4681 agagtggccg ccagcacaca cgtgggagag tctagcctga gcgaagagaa cgacatcttt
     4741 gtgcggacca gcgaggacga gcctgaaagc tctcctcagg acgtggaagt gatcgacgtg
     4801 accgccgatg agatccggct gaaatggtca cctcctgaga agcccaatgg catcattatc
     4861 gcctacgagg tgctgtacaa gaatatcgac accctgtaca tgaagaacac ctccaccacc
     4921 gatatcatcc tgcggaacct gcggcctcac acactgtaca atatcagcgt gcggagctac
     4981 accagatttg gccacggcaa tcaggtgtcc agcctgctga gcgtcagaac ctctgagaca
     5041 gtgcctgata gcgcccctga gaacatcacc tataagaaca tctccagcgg cgagatcgag
     5101 ctgtccttcc tgccaccttc tagccccaac ggaatcatta agaagtatac catctacctg
     5161 aagcggagca acggcaacga ggaacggacc atcaacacca ccagcctgac acagaacatc
     5221 aaggtgctca aaaagtacac gcagtatatc atcgaagtgt ccgcaagcac cctgaaaggc
     5281 gagggcgtta gaagcgcccc tatcagcatt ctgaccgaag aggacgcccc tgacagccca
     5341 cctcaggatt tttccgtgaa gcagctgtcc ggcgtgaccg tgaaactgtc ttggcagcct
     5401 cctctggaac ccaacgggat tatcctgtac tacaccgtgt acgtgtggaa ccgcagcagc
     5461 ctgaaaacca tcaatgtgac cgagacatcc ctggaactga gcgacctgga ctacaacgtg
     5521 gaatactccg cctacgtgac agccagcaca agattcggag atggcaagac ccggtccaac
     5581 atcattagct ttcagacccc tgagggcgct ccctccgatg gcgcgccggg cggcgacaag
     5641 acccacacct gccccccctg cccagctcca gaactgctgg gagggccttc cgtgttcctg
     5701 ttccccccca agcccaagga caccctgatg atcagcagga cccccgaggt gacctgcgtg
     5761 gtggtggccg tgtcccacga ggacccagag gtgaagttca actggtacgt ggacggcgtg
     5821 gaggtgcaca acgccaagac caagcccaga gaggagcagt acaacagcac ctacagggtg
     5881 gtgtccgtgc tgaccgtgct gcaccaggac tggctgaacg gcaaagaata caagtgcaaa
     5941 gtctccaaca aggccctggc tgccccaatc gaaaagacaa tcagcaaggc caagggccag
     6001 ccacgggagc cccaggtgta caccctgccc cccagccggg aggagatgac caagaaccag
     6061 gtgtccctga cctgtctggt gaagggcttc taccccagcg atatcgccgt ggagtgggag
     6121 agcaacggcc agcccgagaa caactacaag accacccccc cagtgctgga cagcgacggc
     6181 agcttcttcc tgtacagcaa gctgaccgtg gacaagtcca ggtggcagca gggcaacgtg
     6241 ttcagctgca gcgtgatgca cgaggccctg cacaaccact acacccagaa gtccctgagc
     6301 ctgagccccg gcggcagcgg tcttaatgat attttcgagg ctcagaagat tgagtggcat
     6361 gaacaccacc accatcacca ctaaggccgg cctgtagtca ctggcacaac gcgttacccg
     6421 gtaagccaat cgggtataca cggtcgtcat actgcagaca gggttcttct actttgcaag
     6481 atagtcttga gtagtaaaat aaatagatag agaaaatcag cctcgactgt gccttctagt
     6541 tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact
     6601 cccactgtcc tttcctaata aaatgaggaa attgcatcac aacactcaac cctatctcgg
     6661 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     6721 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     6781 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     6841 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     6901 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     6961 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     7021 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     7081 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc agcacgtgtt
     7141 gacaattaat catcggcata gtatatcggc atagtataat acgacaaggt gaggaactaa
     7201 accatgaccg agtacaagcc cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc
     7261 gtacgcaccc tcgccgccgc gttcgccgac taccccgcca cgcgccacac cgtcgatccg
     7321 gaccgccaca tcgagcgggt caccgagctg caagaactct tcctcacgcg cgtcgggctc
     7381 gacatcggca aggtgtgggt cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg
     7441 gagagcgtcg aagcgggggc ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc
     7501 ggttcccggc tggccgcgca gcaacagatg gaaggcctcc tggcgccgca ccggcccaag
     7561 gagcccgcgt ggttcctggc caccgtcggc gtctcgcccg accaccaggg caagggtctg
     7621 ggcagcgccg tcgtgctccc cggagtggag gcggccgagc gcgccggggt gcccgccttc
     7681 ctggagacat ccgcgccccg caacctcccc ttctacgagc ggctcggctt caccgtcacc
     7741 gccgacgtcg aggtgcccga aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc
     7801 tgacacgtgc tacgagattt cgattccacc gccgccttct atgaaaggtt gggcttcgga
     7861 atcgttttcc gggacgccgg ctggatgatc ctccagcgcg gggatctcat gctggagttc
     7921 ttcgcccacc ccaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     7981 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     8041 atcaatgtat cttatcatgt ctgtataccg tcgacctcta gctagagctt ggcgtaatca
     8101 tggtcattac caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc
     8161 atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc
     8221 tggccccagc gctgcgatga taccgcgaga accacgctca ccggctccgg atttatcagc
     8281 aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc
     8341 catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt
     8401 gcgcaacgtt gttgccatcg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc
     8461 ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa
     8521 aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt
     8581 atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg
     8641 cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc
     8701 gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa
     8761 agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt
     8821 gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt
     8881 caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag
     8941 ggcgacacgg aaatgttgaa tactcatatt cttcctt
//
