LOCUS       sequence-368532-        6598 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of secreted protein fused to 
            Fc(DAPA)-Avi-6xHis..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6598)
  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 6598)
  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-IL18BP-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6598
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     133..152
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(171..218)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(266..595)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      280..415
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(334..353)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     580..600
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    718..829
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     818..835
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(975..992)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(993..1037)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2346..2365)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2378..2396)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2379..2396)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2606..2630)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2627..2830)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2656..2676)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2831..3210
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3377..3393
                     /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(3849..3866)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3849..3865)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3964..3981)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4135..4723)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4215..4234)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4809..4828)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4834..4853)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4895..4986
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4987..5055,5056..5847)
                     /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(5205..5224)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5900..6021
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5911..5930)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5965..5984
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6151..6598,1..152))
                     /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(6234..6254)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 tcttgcagct cggtgacccg ctcgatgtgg cggtccggat cgacggtgtg gcgcgtggcg
       61 gggtagtcgg cgaacgcggc ggcgagggtg cgtacggccc tggggacgtc gtcgcgggtg
      121 gcgaggcgca ccgtgggctt gtactcggtc atggtttagt tcctcacctt gtcgtattat
      181 actatgccga tatactatgc cgatgattaa ttgtcaacac gtgctgatca gatccgaaaa
      241 tggatataca agctcccggg agctttttgc aaaagcctag gcctccaaaa aagcctcctc
      301 actacttctg gaatagctca gaggcagagg cggcctcggc ctctgcataa ataaaaaaaa
      361 ttagtcagcc atggggcgga gaatgggcgg aactgggcgg agttaggggc gggatgggcg
      421 gagttagggg cgggactatg gttgctgact aattgagatg catgctttgc atacttctgc
      481 ctgctgggga gcctggggac tttccacacc tggttgctga ctaattgaga tgcatgcttt
      541 gcatacttct gcctgctggg gagcctgggg actttccaca ccctaactga cacacattcc
      601 acagaattaa ttcgcgttaa atttttgtta aatcagctca ttttttaacc aataggccga
      661 aatcggcaaa atcccttata aatcaaaaga atagaccgag atagggttga gtgttgtgat
      721 gcaatttcct cattttatta ggaaaggaca gtgggagtgg caccttccag ggtcaaggaa
      781 ggcacggggg aggggcaaac aacagatggc tggcaactag aaggcacagt cgaggctgat
      841 tttctctatc tatttatttt actactcaag actatcttgc aaagtagaag aaccctgtct
      901 gcagtatgac gaccgtgtat acccgattgg cttaccgggt aacgcgttgt gccagtgact
      961 acaggccggc cttagtggtg atggtggtgg tgttcatgcc actcaatctt ctgagcctcg
     1021 aaaatatcat taagaccgct gccgccgggg ctcaggctca gggacttctg ggtgtagtgg
     1081 ttgtgcaggg cctcgtgcat cacgctgcag ctgaacacgt tgccctgctg ccacctggac
     1141 ttgtccacgg tcagcttgct gtacaggaag aagctgccgt cgctgtccag cactgggggg
     1201 gtggtcttgt agttgttctc gggctggccg ttgctctccc actccacggc gatatcgctg
     1261 gggtagaagc ccttcaccag acaggtcagg gacacctggt tcttggtcat ctcctcccgg
     1321 ctggggggca gggtgtacac ctggggctcc cgtggctggc ccttggcctt gctgattgtc
     1381 ttttcgattg gggcagccag ggccttgttg gagactttgc acttgtattc tttgccgttc
     1441 agccagtcct ggtgcagcac ggtcagcacg gacaccaccc tgtaggtgct gttgtactgc
     1501 tcctctctgg gcttggtctt ggcgttgtgc acctccacgc cgtccacgta ccagttgaac
     1561 ttcacctctg ggtcctcgtg ggacacggcc accaccacgc aggtcacctc gggggtcctg
     1621 ctgatcatca gggtgtcctt gggcttgggg gggaacagga acacggaagg ccctcccagc
     1681 agttctggag ctgggcaggg ggggcaggtg tgggtcttgt cgccgcccgg cgcgccacct
     1741 tgttgctgag gactgctatg agagctaggc agggcctctt gtgtaggagg cagtgtagct
     1801 ctcagtccgg cccacagttg agccagaacc acatgtctct gcaccacctg ttcagggtcc
     1861 accagcacgc agctgaaatt ggtgctgtgc agagctggtg tcagttgttc cagcaccagg
     1921 gctttgcaca gctgggtgcc tgtagatcct ctctctctgc ttgtgctgcc ctcccacagt
     1981 ctaccaggca gatgttcgat gaagctgccg ttgccgagcc agtacaggat gctgaagttg
     2041 gggaatctgc tgcaggccac acagctcagg ctcagtgtgc cattcagagg cacttccact
     2101 tcgggccatg tcacttccag agcgggacac tgtttagcgg cagggaacac aggaggctgg
     2161 ctaggacaag gatcttttgt gctccgcacg ctggctgtgg cggctgttgt tgtttgagac
     2221 acaggtgtgg cggccgcagc gaacaccagg cagaagatgt agctcagggc aatgatggtc
     2281 ttcatggtgg cgctagcagg cgcgttttcg atagaaccga ggtgcagttg gacctgggag
     2341 tggacacctg tggagagaaa ggcaaagtgg atgttattct atagtgtcac ctaaatcgta
     2401 tgtgtatgat acataaggtt atgtattaat tgtagccgcg ttctaacgaa gccaaggggg
     2461 tgggcctata gactctatag gcggtactta cgtcactctt ggcacgggga atccgcgttc
     2521 caatgcaccg ttcccggccg cggaggctgg atcggtcccg gtgtcttcta tggaggtcaa
     2581 aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct ctgcttatat
     2641 agacctccca ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac
     2701 attttggaaa gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt
     2761 ggagacttgg aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa
     2821 accgcatcac catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt
     2881 cccataaggt catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa
     2941 tagggggcgt acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta
     3001 aatactccac ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg
     3061 tcattattga cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg
     3121 taagttatgt aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt
     3181 actattaata actagtcaat aatcaatgcc aacatggcgg tcatattgga catgagccaa
     3241 tataaatgta catattatga tatagataca acgtatgcaa tggccaatag ccaatattga
     3301 tttatgctat ataaccaatg aataatatgg ctaatggcca atattgagtc aacgccactg
     3361 tgcatgaagc tcgtaattgt tatccgctca caattaaggg cgacacaaaa tttattctaa
     3421 atgataataa atactgataa catcttatag tttgtattat attttgtatt atcgttgaca
     3481 tgtataattt tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt
     3541 aattctcttt aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat
     3601 agtttaatta taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc
     3661 ttttttctca tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct
     3721 taaatattct gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg
     3781 ggtttttacg ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg
     3841 agcttaagac tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc
     3901 atccgtcagg ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc
     3961 ttcctcgctc actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca
     4021 ctcaaaggcg gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg
     4081 agcaaaaggc cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca
     4141 taggctccgc ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa
     4201 cccgacagga ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc
     4261 tgttccgacc ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc
     4321 gctttctcat agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct
     4381 gggctgtgtg cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg
     4441 tcttgagtcc aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag
     4501 gattagcaga gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta
     4561 cggctacact agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg
     4621 aaaaagagtt ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt
     4681 tgtttgcaag cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt
     4741 ttctacgggg tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg
     4801 gtcatgagca aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc
     4861 agaattggtt aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca
     4921 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     4981 aagaatatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     5041 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     5101 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     5161 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     5221 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     5281 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     5341 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     5401 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     5461 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     5521 acgatgcctg tagcgatggc aacaacgttg cgcaaactat taactggcga actacttact
     5581 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     5641 ctgcgctcgg cccttccggc tggctggttt attgctgata aatccggagc cggtgagcgt
     5701 ggttctcgcg gtatcatcgc agcgctgggg ccagatggta agccctcccg tatcgtagtt
     5761 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     5821 ggtgcctcac tgattaagca ttggtaatga ccatgattac gccaagctct agctagaggt
     5881 cgacggtata cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     5941 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt
     6001 ataagctgca ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg
     6061 gaggatcatc cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc
     6121 ggcggtggaa tcgaaatctc gtagcacgtg tcaggcaccg ggcttgcggg tcatgcacca
     6181 ggtgcgcggt ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta
     6241 gaaggggagg ttgcggggcg cggatgtctc caggaaggcg ggcaccccgg cgcgctcggc
     6301 cgcctccact ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga
     6361 gacgccgacg gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc
     6421 ttccatctgt tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat
     6481 ctcggcgaac accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc
     6541 gccgtcgtcc gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagt
//
