LOCUS       sequence-447075-        8731 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 8731)
  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 8731)
  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-SIGLEC1(Trunc2)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8731
                     /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             6073..6117
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6118..6135
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6275..6292)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6281..6392
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6510..6530)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6515..6844
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6695..6830
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6757..6776
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6892..6939
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6958..7557
                     /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(6958..6977)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7454..7474
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7687..7808
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7724..7743)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7778..7797
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7861..8652,8653..8721))
                     /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     8484..8503
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8722..8731,1..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 cggccgcctc ttggggagtt
     2761 agttctcctc aggatgtgca gggcgtgaag ggctcttgtc tgctgatccc ctgcatcttc
     2821 agcttccccg ctgacgtgga agtgcctgat ggaatcaccg ccatctggta ctacgactac
     2881 agcggccaga gacaggtggt gtctcactct gccgatccta agctggtgga agccagattc
     2941 agaggcagaa ccgagttcat gggcaacccc gagcacagag tgtgcaacct gctgctgaaa
     3001 gacctgcagc ctgaggacag cggcagctac aacttcagat tcgagatcag cgaagtgaac
     3061 cgttggagcg acgtgaaggg aaccctggtc acagtgaccg aggaacccag agtgcctaca
     3121 atcgcctctc ctgtggaact gctggaaggc accgaggtgg acttcaactg tagcacccct
     3181 tacgtgtgcc tgcaagagca agttcggctg cagtggcagg gacaagatcc tgccagaagc
     3241 gtgaccttca acagccagaa gtttgagcca acaggcgtgg gccacctgga aaccctgcat
     3301 atggctatgt cttggcagga ccacggcaga atcctgagat gccagctgag cgtggccaat
     3361 cacagagccc agagcgagat ccatctgcaa gtgaagtatg cccctaaagg cgtgaagatt
     3421 ctgctgagcc ccagcggcag aaatatcctg cctggcgagc tggtcaccct gacctgtcaa
     3481 gtgaatagca gctaccccgc cgtgtccagc atcaagtggc tgaaggatgg cgtgcggctg
     3541 cagacaaaaa ctggcgttct gcatctgcct caggccgctt ggagtgatgc tggcgtgtac
     3601 acctgtcagg ccgagaatgg cgttggctct ctggtgtctc ctcctatctc tctgcacatc
     3661 ttcatggccg aggtccaggt ttcccctgcc ggacctatcc tggaaaacca gaccgtgaca
     3721 ctcgtgtgca acacccctaa tgaggcccct agcgacctgc ggtacagctg gtataagaac
     3781 cacgtgctgc tcgaggacgc ccactctcac acactgagac tgcatctggc caccagagcc
     3841 gataccggct tctacttctg cgaggtgcag aatgtgcacg gctctgagag atctggccct
     3901 gtgtccgtgg tggtcaatca ccctccactg acacctgtgc tgaccgcctt tctggaaaca
     3961 caggccggac tcgtgggcat cctgcactgt tctgtggtgt ctgaacctct ggccacactg
     4021 gtgctgtctc acggcggaca tattctggcc agcacaagcg gcgatagcga tcacagccct
     4081 agattcagcg gcaccagcgg acctaacagc ctgcggctgg aaatccgcga cctggaagaa
     4141 acagactccg gcgagtacaa gtgcagcgcc accaattctc tgggcaacgc caccagcaca
     4201 ctggactttc atgccaatgc cgccagactg ctgatttctc ctgctgccga agtggtggaa
     4261 ggacaggctg tgacactgtc ctgtagaagc ggactgagcc ctacacctga tgccaggttt
     4321 agctggtatc tgaacggcgc cctgctgcat gaaggccctg gatcttctct tctgctgcct
     4381 gccgcttcta gcaccgatgc cggctcttat cactgtagag ccagagatgg acacagcgcc
     4441 tctggaccat cttctccagc tgtcctgaca gtgctgtacc ctcctagaca gcccaccttc
     4501 accaccagac tggatttgga tgctgctggt gctggcgctg ggagaagagg actgctgctg
     4561 tgtagagtgg acagcgatcc tcctgctaga ctgcagctgc tgcacaagga tagagtggtg
     4621 gccacatctc tgcctagcgg cggaggatgt tctacctgtg gcggctgtag ccccagaatg
     4681 aaggttacaa aggcccctaa cctgctgagg gtcgagatcc acaatcctct gctcgaagag
     4741 gaaggcctgt acctctgcga agccagcaat gccctgggca atgccagcac ctctgccacc
     4801 tttaatggac aggccaccgt gctggctatc gcccctagtc atacactgca agagggcaca
     4861 gaggccaacc tgacctgcaa tgtgtcaaga gaagccgccg gatctcccgc caacttcagc
     4921 tggtttagaa acggcgtgct gtgggcccag ggacctcttg aaacagtgac actgctgccc
     4981 gtggccagaa cagatgccgc actgtatgcc tgtcggattc tgacagaagc tggcgcccag
     5041 ctgtctacac ctgttctgct gtctgtgctg tatccacctg acagacctaa gctgtctgcc
     5101 ctgctggata tgggacaggg acacatggcc ctgttcatct gcaccgtgga tagcagacca
     5161 ctggctctgc ttgccctgtt tcatggcgaa catctgctgg ccacaagcct gggacctcag
     5221 gtgccatctc atggcagatt tcaggccaag gccgaggcca atagcctgaa gctggaagtc
     5281 cgagaactcg gcctgggcga ttctggcagc tatagatgcg aggccacaaa cgtgctgggc
     5341 agcagcaaca ccagcctgtt ctttcaagtc cgaggcgcgc cgggcggcga caagacccac
     5401 acctgccccc cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc
     5461 cccaagccca aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg
     5521 gccgtgtccc acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg
     5581 cacaacgcca agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc
     5641 gtgctgaccg tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc
     5701 aacaaggccc tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg
     5761 gagccccagg tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc
     5821 ctgacctgtc tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac
     5881 ggccagcccg agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc
     5941 ttcctgtaca gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc
     6001 tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc
     6061 cccggcggca gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac
     6121 caccaccatc accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc
     6181 caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc
     6241 ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag
     6301 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     6361 gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt
     6421 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     6481 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     6541 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     6601 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6661 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     6721 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     6781 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     6841 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     6901 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     6961 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     7021 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     7081 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     7141 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     7201 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     7261 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     7321 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     7381 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     7441 acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     7501 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac
     7561 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     7621 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     7681 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     7741 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     7801 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     7861 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     7921 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7981 cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa
     8041 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     8101 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     8161 cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     8221 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     8281 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     8341 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     8401 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     8461 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     8521 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     8581 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     8641 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     8701 acggaaatgt tgaatactca tattcttcct t
//
