LOCUS       sequence-368511-        6553 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein 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 6553)
  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 6553)
  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-PDCD1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6553
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     353..372
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(391..438)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(486..815)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      500..635
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(554..573)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     800..820
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    938..1049
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1038..1055
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1195..1212)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1213..1257)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2521..2540)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2553..2571)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2554..2571)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2781..2805)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2802..3005)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2831..2851)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        3006..3385
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3552..3568
                     /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)."
     primer_bind     complement(4024..4041)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(4024..4040)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4139..4156)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4310..4898)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4390..4409)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4984..5003)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(5009..5028)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        5070..5161
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5162..5230,5231..6022)
                     /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     complement(5380..5399)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    6075..6196
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6086..6105)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     6140..6159
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6326..6553,1..372))
                     /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(6409..6429)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gaaccacgcg ggctccttgg gccggtgcgg cgccaggagg ccttccatct gttgctgcgc
       61 ggccagccgg gaaccgctca actcggccat gcgcgggccg atctcggcga acaccgcccc
      121 cgcttcgacg ctctccggcg tggtccagac cgccaccgcg gcgccgtcgt ccgcgaccca
      181 caccttgccg atgtcgagcc cgacgcgcgt gaggaagagt tcttgcagct cggtgacccg
      241 ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc
      301 ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt
      361 gtactcggtc atggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc
      421 cgatgattaa ttgtcaacac gtgctgatca gatccgaaaa tggatataca agctcccggg
      481 agctttttgc aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca
      541 gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga
      601 gaatgggcgg aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg
      661 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      721 tttccacacc tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg
      781 gagcctgggg actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa
      841 atttttgtta aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata
      901 aatcaaaaga atagaccgag atagggttga gtgttgtgat gcaatttcct cattttatta
      961 ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac
     1021 aacagatggc tggcaactag aaggcacagt cgaggctgat tttctctatc tatttatttt
     1081 actactcaag actatcttgc aaagtagaag aaccctgtct gcagtatgac gaccgtgtat
     1141 acccgattgg cttaccgggt aacgcgttgt gccagtgact acaggccggc cttagtggtg
     1201 atggtggtgg tgttcatgcc actcaatctt ctgagcctcg aaaatatcat taagaccgct
     1261 gccgccgggg ctcaggctca gggacttctg ggtgtagtgg ttgtgcaggg cctcgtgcat
     1321 cacgctgcag ctgaacacgt tgccctgctg ccacctggac ttgtccacgg tcagcttgct
     1381 gtacaggaag aagctgccgt cgctgtccag cactgggggg gtggtcttgt agttgttctc
     1441 gggctggccg ttgctctccc actccacggc gatatcgctg gggtagaagc ccttcaccag
     1501 acaggtcagg gacacctggt tcttggtcat ctcctcccgg ctggggggca gggtgtacac
     1561 ctggggctcc cgtggctggc ccttggcctt gctgattgtc ttttcgattg gggcagccag
     1621 ggccttgttg gagactttgc acttgtattc tttgccgttc agccagtcct ggtgcagcac
     1681 ggtcagcacg gacaccaccc tgtaggtgct gttgtactgc tcctctctgg gcttggtctt
     1741 ggcgttgtgc acctccacgc cgtccacgta ccagttgaac ttcacctctg ggtcctcgtg
     1801 ggacacggcc accaccacgc aggtcacctc gggggtcctg ctgatcatca gggtgtcctt
     1861 gggcttgggg gggaacagga acacggaagg ccctcccagc agttctggag ctgggcaggg
     1921 ggggcaggtg tgggtcttgt cgccgcccgg cgcgccaacg agggtctgaa actggccggc
     1981 aggtcttgga gatggagatg ggtgagctgt gggcacttca gcccgtcttt cggtcactct
     2041 cagctcggct ctcaggctct ctttgatctg agccttaggg gccagagaaa tggcgccgca
     2101 cagatatgtg ccgctgtcgt ttcttctggc ccgcacaaca gacatgtgga agtctctgcc
     2161 gttgggcagc tgtgtcactc tgaaccggca gtcctgtcca ggctggcttc tatcctcagg
     2221 aaaggcggcc agcttgtcgg tctggttgct ggggctcatt ctgtaccagt tcagcacgaa
     2281 gctctcgctg gtgttgctga agctacaggt gaaggtggca ttatcgccct ctgtgaccac
     2341 cagcagagcg gggctgaatg taggaggatt ccagggtcta tcgggagaat ccagaaacca
     2401 tccggcggcc gcagcgaaca ccaggcagaa gatgtagctc agggcaatga tggtcttcat
     2461 ggtggcgcta gcaggcgcgt tttcgataga accgaggtgc agttggacct gggagtggac
     2521 acctgtggag agaaaggcaa agtggatgtt attctatagt gtcacctaaa tcgtatgtgt
     2581 atgatacata aggttatgta ttaattgtag ccgcgttcta acgaagccaa gggggtgggc
     2641 ctatagactc tataggcggt acttacgtca ctcttggcac ggggaatccg cgttccaatg
     2701 caccgttccc ggccgcggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag
     2761 cgtggatggc gtctccaggc gatctgacgg ttcactaaac gagctctgct tatatagacc
     2821 tcccaccgta cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt
     2881 ggaaagtccc gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga
     2941 cttggaaatc cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc
     3001 atcaccatgg taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat
     3061 aaggtcatgt actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg
     3121 ggcgtacttg gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac
     3181 tccacccatt gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt
     3241 attgacgtca atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt
     3301 tatgtaacgc ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat
     3361 taataactag tcaataatca atgccaacat ggcggtcata ttggacatga gccaatataa
     3421 atgtacatat tatgatatag atacaacgta tgcaatggcc aatagccaat attgatttat
     3481 gctatataac caatgaataa tatggctaat ggccaatatt gagtcaacgc cactgtgcat
     3541 gaagctcgta attgttatcc gctcacaatt aagggcgaca caaaatttat tctaaatgat
     3601 aataaatact gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat
     3661 aattttgata tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc
     3721 tctttaacaa actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt
     3781 aattataggt gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt
     3841 tctcatttat aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat
     3901 attctgacaa atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt
     3961 ttacgttatt tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt
     4021 aagactggcc gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg
     4081 tcaggggcct tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct
     4141 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4201 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4261 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4321 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4381 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4441 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4501 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4561 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4621 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4681 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct
     4741 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4801 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4861 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     4921 cggggtctga cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat
     4981 gagcaagggg tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat
     5041 tggttaattg gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat
     5101 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa
     5161 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5221 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5281 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5341 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5401 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5461 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5521 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     5581 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     5641 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     5701 gcctgtagcg atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     5761 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     5821 ctcggccctt ccggctggct ggtttattgc tgataaatcc ggagccggtg agcgtggttc
     5881 tcgcggtatc atcgcagcgc tggggccaga tggtaagccc tcccgtatcg tagttatcta
     5941 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6001 ctcactgatt aagcattggt aatgaccatg attacgccaa gctctagcta gaggtcgacg
     6061 gtatacagac atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg
     6121 aaaaaaatgc tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag
     6181 ctgcaataaa caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga
     6241 tcatccagcc ggcgtcccgg aaaacgattc cgaagcccaa cctttcatag aaggcggcgg
     6301 tggaatcgaa atctcgtagc acgtgtcagg caccgggctt gcgggtcatg caccaggtgc
     6361 gcggtccttc gggcacctcg acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg
     6421 ggaggttgcg gggcgcggat gtctccagga aggcgggcac cccggcgcgc tcggccgcct
     6481 ccactccggg gagcacgacg gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc
     6541 cgacggtggc cag
//
