LOCUS       sequence-303159-        9214 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 9214)
  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 9214)
  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-SDK2(Trunc3)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..9214
                     /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             6556..6600
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6601..6618
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6758..6775)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6764..6875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6993..7013)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6998..7327
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7178..7313
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7240..7259
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7375..7422
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7441..8040
                     /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(7441..7460)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7937..7957
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8170..8291
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8207..8226)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8261..8280
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(8344..9135,9136..9204))
                     /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     8967..8986
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(9205..9214,1..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 tgttggacat
     2761 ctgagcttta gcgagatcct ggacaccagc ctgaaggtgt cctggcaaga gcccggcgag
     2821 aagaatggca tcctgaccgg ctaccggatc agctgggaag agtacaaccg gaccaacacc
     2881 agagtgaccc actacctgcc taacgtgacc ctcgagtaca gagtgacagg cctgacagcc
     2941 ctgaccacct acacaattga agtggccgcc atgaccagca aaggccaggg acaagtgtct
     3001 gccagcacaa tctctagcgg cgtgccacct gaactgcctg gacctcctac aaacctgggc
     3061 atcagcaaca tcggccccag aagcgtgacc ctgcagttca gaccaggcta cgatggcaag
     3121 accagcatct ctcggtggct ggtggaagcc caagtgggcg ttgtcggaga aggcgaggaa
     3181 tggctgctga tccaccagct gagcaacgag cctgacgcca gatccatgga agtgcccgat
     3241 ctgaacccct tcacctgtta cagcttccgg atgcggcaag tgaacatcgt gggcacaagc
     3301 cctccaagcc agcctagcag aaagatccag acactgcagg cccctccaga tatggcccct
     3361 gccaatgtgt ctctgagaac cgccagcgag acaagcctgt ggctgagatg gatgcccctg
     3421 cctgaaatgg agtacaacgg caaccctgag agcgtgggct acaagatcaa gtacagcaga
     3481 agcgacggcc acggcaagac actgtctcac gtggtgcagg acagagtgga acgggactac
     3541 accatcgagg acctggaaga gtggaccgag tatagagtcc aggtgcaggc cttcaacgcc
     3601 atcggatctg gaccttggag ccagacagtc gtgggcagaa ccagagaaag cgtgccaagc
     3661 agcggcccca caaatgtgtc tgccctggcc acaacaagca gcagcatgct cgttcgttgg
     3721 agcgaagtgc ctgaggccga cagaaatgga ctggtgctgg gatacaaagt gatgtacaaa
     3781 gagaaggaca gcgacaccca gcctcggttc tggctggttg agggcaatag ctccagaagc
     3841 gcccagctga caggactggg caaatacgtg ctgtacgagg tgcaggtcct ggccttcaca
     3901 agaatcggag atggcagccc cagccatcct cctatcctgg aaagaaccct ggacgacgtg
     3961 cccggacctc caatgggaat tctgttccct gaagtgcgga ccaccagcgt gcgactgatt
     4021 tggcaacctc ctgccgctcc taacggcatc atcctggcct accagatcac ccacagactg
     4081 aacaccacca ccgccaatac cgccaccgtg gaagttctgg cccctagcgc aagacagtac
     4141 accgccacag gactgaagcc cgagagcgtg tacctgttca gaatcacagc ccagaccaga
     4201 aaaggctggg gcgaagctgc tgaagccctg gtggttacca ccgagaagag agacagaccc
     4261 cagcctccta gcagacccat ggtgcagcaa gaagatgtgc gggccagatc cgtgctgctg
     4321 tcttgggaac ctggctccga tggactgagc cctgtgcggt actacacaat ccagacaaga
     4381 gagctgccca gcggcagatg ggctctgcat tctgcctccg tgtctcacaa cgcctccagc
     4441 ttcatcgtgg accggctgaa gcctttcacc agctacaagt ttagagtcaa ggccaccaac
     4501 gacatcggcg acagcgagtt cagcgaggaa agcgagagcc tgaccactct gcaggctgcc
     4561 cctgatgagg cccctacaat cctgtctgtg acccctcaca ccacaacctc cgtgctgatt
     4621 aggtggcagc ctccggccga ggataagatc aacggaatcc tgctgggctt cagaatccgg
     4681 tacagagaac tgctgtatga gggcctgaga ggcttcaccc tgcggggcat taacaatcct
     4741 ggcgccacat gggccgagct gaccagcatg tacagcatgc ggaacctgag caggcccagc
     4801 ctgacacagt acgagctgga caacctgaac aaacatcgga gatacgagat ccggatgtcc
     4861 gtgtacaatg ccgttggcga gggacctagc agccctcctc aagaagtgtt tgtcggcgag
     4921 gccgtgccta cagctgcccc tagaaatgtg gtggtgcacg gcgctacagc cacacagctg
     4981 gatgtgacat gggagcctcc acctctggac agccagaatg gcgatatcca gggctataag
     5041 atctacttct gggaagccca gagaggcaac ctgaccgaga gagtgaaaac cctgttcctg
     5101 gccgagaaca gcgtgaagct gaagaacctg actggctaca ccgcctacat ggtgtccgtg
     5161 gccgccttta atgctgctgg cgacggccct agaagcaccc ctacacaggg acaaacacag
     5221 caggctgctc ctagcgctcc aagctccgtg aagttctccg aactgaccac cacaagcgtc
     5281 aacgtgtcct gggaagctcc tcagttccca aacggcatcc tggaaggcta cagactggtg
     5341 tacgagcctt gcagccctgt ggatggcgtg tccaagatcg tgaccgtgga cgtgaagggc
     5401 aacagccctc tgtggctcaa agtgaaggat ctggctgagg gcgtgaccta ccggttcagg
     5461 attagagcca agaccttcac atacggcccc gagatcgagg ccaacgtgac aacaggacct
     5521 ggcgaaggtg ctccaggacc tcctggcgtg ccaatcatcg tgcggtacag ctctgccatt
     5581 gccatccact ggtcctctgg cgatcctggc aaaggcccca tcaccagata cgtgatcgag
     5641 gctagaccca gcgacgaagg cctgtgggac atcctgatca aggacatccc caaagaggtg
     5701 tccagctaca ccttctccat ggatatcctg aagccaggcg tgtcctacga cttcagagtg
     5761 atcgccgtga acgactacgg cttcggcaca ccttctagcc ctagccagtc tgtgccagct
     5821 cagaaggcca atcctttcta cgaagagtgg tggtttggcg cgccgggcgg cgacaagacc
     5881 cacacctgcc ccccctgccc agctccagaa ctgctgggag ggccttccgt gttcctgttc
     5941 ccccccaagc ccaaggacac cctgatgatc agcaggaccc ccgaggtgac ctgcgtggtg
     6001 gtggccgtgt cccacgagga cccagaggtg aagttcaact ggtacgtgga cggcgtggag
     6061 gtgcacaacg ccaagaccaa gcccagagag gagcagtaca acagcaccta cagggtggtg
     6121 tccgtgctga ccgtgctgca ccaggactgg ctgaacggca aagaatacaa gtgcaaagtc
     6181 tccaacaagg ccctggctgc cccaatcgaa aagacaatca gcaaggccaa gggccagcca
     6241 cgggagcccc aggtgtacac cctgcccccc agccgggagg agatgaccaa gaaccaggtg
     6301 tccctgacct gtctggtgaa gggcttctac cccagcgata tcgccgtgga gtgggagagc
     6361 aacggccagc ccgagaacaa ctacaagacc acccccccag tgctggacag cgacggcagc
     6421 ttcttcctgt acagcaagct gaccgtggac aagtccaggt ggcagcaggg caacgtgttc
     6481 agctgcagcg tgatgcacga ggccctgcac aaccactaca cccagaagtc cctgagcctg
     6541 agccccggcg gcagcggtct taatgatatt ttcgaggctc agaagattga gtggcatgaa
     6601 caccaccacc atcaccacta aggccggcct gtagtcactg gcacaacgcg ttacccggta
     6661 agccaatcgg gtatacacgg tcgtcatact gcagacaggg ttcttctact ttgcaagata
     6721 gtcttgagta gtaaaataaa tagatagaga aaatcagcct cgactgtgcc ttctagttgc
     6781 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     6841 actgtccttt cctaataaaa tgaggaaatt gcatcacaac actcaaccct atctcggtct
     6901 attcttttga tttataaggg attttgccga tttcggccta ttggttaaaa aatgagctga
     6961 tttaacaaaa atttaacgcg aattaattct gtggaatgtg tgtcagttag ggtgtggaaa
     7021 gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac
     7081 caggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     7141 ttagtcagca accatagtcc cgcccctaac tccgcccatc ccgcccctaa ctccgcccag
     7201 ttccgcccat tctccgcccc atggctgact aatttttttt atttatgcag aggccgaggc
     7261 cgcctctgcc tctgagctat tccagaagta gtgaggaggc ttttttggag gcctaggctt
     7321 ttgcaaaaag ctcccgggag cttgtatatc cattttcgga tctgatcagc acgtgttgac
     7381 aattaatcat cggcatagta tatcggcata gtataatacg acaaggtgag gaactaaacc
     7441 atgaccgagt acaagcccac ggtgcgcctc gccacccgcg acgacgtccc cagggccgta
     7501 cgcaccctcg ccgccgcgtt cgccgactac cccgccacgc gccacaccgt cgatccggac
     7561 cgccacatcg agcgggtcac cgagctgcaa gaactcttcc tcacgcgcgt cgggctcgac
     7621 atcggcaagg tgtgggtcgc ggacgacggc gccgcggtgg cggtctggac cacgccggag
     7681 agcgtcgaag cgggggcggt gttcgccgag atcggcccgc gcatggccga gttgagcggt
     7741 tcccggctgg ccgcgcagca acagatggaa ggcctcctgg cgccgcaccg gcccaaggag
     7801 cccgcgtggt tcctggccac cgtcggcgtc tcgcccgacc accagggcaa gggtctgggc
     7861 agcgccgtcg tgctccccgg agtggaggcg gccgagcgcg ccggggtgcc cgccttcctg
     7921 gagacatccg cgccccgcaa cctccccttc tacgagcggc tcggcttcac cgtcaccgcc
     7981 gacgtcgagg tgcccgaagg accgcgcacc tggtgcatga cccgcaagcc cggtgcctga
     8041 cacgtgctac gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc
     8101 gttttccggg acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc
     8161 gcccacccca acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca
     8221 aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc
     8281 aatgtatctt atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg
     8341 tcattaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc
     8401 catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg
     8461 ccccagcgct gcgatgatac cgcgagaacc acgctcaccg gctccggatt tatcagcaat
     8521 aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat
     8581 ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg
     8641 caacgttgtt gccatcgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc
     8701 attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa
     8761 agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc
     8821 actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt
     8881 ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag
     8941 ttgctcttgc ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt
     9001 gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag
     9061 atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac
     9121 cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc
     9181 gacacggaaa tgttgaatac tcatattctt cctt
//
