LOCUS       sequence-303153-        9163 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 9163)
  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 9163)
  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-SDK1(Trunc3)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..9163
                     /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             6505..6549
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6550..6567
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6707..6724)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6713..6824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6942..6962)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6947..7276
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7127..7262
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7189..7208
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7324..7371
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7390..7989
                     /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(7390..7409)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7886..7906
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8119..8240
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8156..8175)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8210..8229
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8293..9084,9085..9153))
                     /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     8916..8935
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9154..9163,1..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 cggccgccgc tgttggacac
     2761 ctgagcttta cagagatcct ggacaccagc ctgaaggtgt cctggcaaga gcccctggaa
     2821 aagaacggca tcatcaccgg ctaccagatc agctgggaag tgtacggcag aaacgacagc
     2881 agactgaccc acacactgaa cagcaccaca cacgagtaca agatccaggg cctgagcagc
     2941 ctgaccacct acacaattga tgtggccgcc gtgacagccg ttggcacagg actggtcaca
     3001 agcagcacaa tctctagcgg cgtgccacct gatttgcctg gcgctcctag caacctggtc
     3061 atcagcaaca tcagccccag aagcgccaca ctgcagttca gaccaggcta cgatggcaag
     3121 accagcatca gcagatggat cgtggaaggc caagtgggag ccatcggaga tgaggaagag
     3181 tgggtcaccc tgtacgagga agagaacgag cctgacgctc agatgctgga aatccccaat
     3241 ctgacccctt acacacacta ccggttccgg atgaagcaag tgaacatcgt gggcccatct
     3301 ccatacagcc ccagcagcag agtgattcag acactgcagg ctcctccaga tgtggcccct
     3361 acaagcgtga cagtgcggac agcctctgag acaagcctga gactgagatg ggtgcccctg
     3421 cctgatagcc agtacaacgg caatcctgag agcgtgggct acagaatcaa gtactggcgg
     3481 agcgatctgc agtctagcgc tgtggctcag gtggtgtctg accggctgga aagagagttc
     3541 accatcgagg aactggaaga atggatggaa tacgagctgc agatgcaggc cttcaatgcc
     3601 gttggagctg gaccttggag cgaggttgtc agaggcagaa ccagagaaag cgtgccatct
     3661 gccgctccag agaatgtgtc tgccgaagcc gtgtctagca cccagatcct gctgacctgg
     3721 acatccgtgc cagagcagga tcagaatggc ctgatcctgg gctataagat cctgttcaga
     3781 gccaaggatc tggaccccga gcctagatct cacatcgtgc ggggcaatca cacacagtct
     3841 gctctgctgg ccggactgag aaagttcgtg ctgtatgagc tgcaggtcct ggcctttacc
     3901 agaatcggaa atggcgtgcc cagcacaccc ctgattctgg aaagaacaaa ggacgacgcc
     3961 cctggacctc ctgtcagact ggtttttcct gaagtgcggc tgaccagcgt gcggattgtg
     4021 tggcaacctc ctgaggaacc caacggaatc atcctgggat accagatcgc ctacagactg
     4081 gccagcagct cccctcacac ctttacaaca gtggaagtgg gcgccaccgt gcggcagttt
     4141 acagctacag atctggctcc cgagagcgcc tacatcttca gactgagcgc caagacaaga
     4201 caaggctggg gagaacctct ggaagccacc gtgatcacca ccgagaagag agaaagaccc
     4261 gctcctccta gagaactgct ggtgcctcag gccgaagtga ccgctagatc tctgagactt
     4321 cagtgggtgc caggctctga tggcgcctct cctatccggt acttcaccat gcaagtgcgc
     4381 gagctgccta gaggcgagtg gcagacatac agcagctcta tctctcacga ggccaccgcc
     4441 tgtgtggtgg atagactgag gcctttcacc agctacaagc tgcggctgaa ggccaccaac
     4501 gacatcggcg atagcgactt cagcagcgag acagaggcag tgacaaccct gcaggatgtt
     4561 cctggcgagc ctcctggatc tgtgtccgcc acacctcaca caacaagcag cgttctgatc
     4621 cagtggcagc ctcctaggga tgagagcctg aatggactgc tgcagggcta tagaatctac
     4681 taccgcgagc tggaatatga ggccggcagc ggaacagagg ccaagacact gaagaaccct
     4741 atcgctctgc acgccgagct gacagcccag agcagcttta agaccgtgaa cagcagcagc
     4801 acctccacca tgtgcgagct gacccacctg aagaagtatc ggagatacga agtgatcatg
     4861 accgcctaca acatcatcgg cgagagccct gcttctgccc ctgtggaagt gtttgtggga
     4921 gaagccgctc ctgccatggc tccccagaat gtgcaagtga cacctctgac agccagccag
     4981 ctggaagtta catgggaccc tccaccacca gagagccaga acggcaatat tcagggctac
     5041 aagatctact attgggaagc cgacagccag aatgagacag agaagatgaa ggtgctgttt
     5101 ctgcccgagc ctgtcgtgcg cctgaagaat ctgaccagcc acacaaagta cctggtgtct
     5161 atcagcgcct tcaacgctgc tggcgacggc cctaaatctg atcctcagca gggaagaaca
     5221 caccaggcag ctccaggcgc tccaagcttt ctggcctttt ccgagatcac cagcaccact
     5281 ctcaatgtgt cttggggaga gcctgccgcc gctaatggca ttctgcaggg atacagagtg
     5341 gtgtacgagc ctctggctcc agtgcagggc gtgtccaaag tggtcaccgt ggaagttcgc
     5401 ggcaactggc agagatggct gaaagtgcgg gatctgacca agggcgtgac ctacttcttt
     5461 cgggtgcaag ccagaaccat cacatacgga cccgaactgc aggccaatat cacagctgga
     5521 ccagccgaag gatctccagg cagccctaga gatgtcctcg tgacaaagag cgccagcgaa
     5581 ctgaccctgc agtggacaga aggacacagc ggcgataccc ctaccacagg ctatgtgatc
     5641 gaggccagac cttctgacga aggcctgtgg gacatgttcg tgaaggacat ccctcggagc
     5701 gccacaagct acaccctgag cctggataag ctgagacagg gcgtcacata cgagtttaga
     5761 gtggtggccg tgaacgaggc cggctacggc gaaccttcta atcctggcgc gccgggcggc
     5821 gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg
     5881 ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc
     5941 tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac
     6001 ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac
     6061 agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag
     6121 tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag
     6181 ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag
     6241 aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag
     6301 tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc
     6361 gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcagggc
     6421 aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc
     6481 ctgagcctga gccccggcgg cagcggtctt aatgatattt tcgaggctca gaagattgag
     6541 tggcatgaac accaccacca tcaccactaa ggccggcctg tagtcactgg cacaacgcgt
     6601 tacccggtaa gccaatcggg tatacacggt cgtcatactg cagacagggt tcttctactt
     6661 tgcaagatag tcttgagtag taaaataaat agatagagaa aatcagcctc gactgtgcct
     6721 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     6781 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcacaaca ctcaacccta
     6841 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6901 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     6961 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     7021 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     7081 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     7141 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     7201 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     7261 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     7321 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     7381 aactaaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     7441 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     7501 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     7561 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     7621 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     7681 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     7741 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     7801 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7861 gccttcctgg agacatccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7921 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7981 ggtgcctgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     8041 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     8101 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     8161 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     8221 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     8281 taatcatggt cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     8341 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     8401 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
     8461 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     8521 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     8581 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     8641 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     8701 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     8761 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     8821 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8881 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8941 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     9001 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     9061 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     9121 aataagggcg acacggaaat gttgaatact catattcttc ctt
//
