LOCUS       sequence-420967-        6670 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 6670)
  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 6670)
  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-ULBP1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6670
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     51..70
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(89..136)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(184..513)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      198..333
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(252..271)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     498..518
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    636..747
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     736..753
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(893..910)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(911..955)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2336..2355)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2368..2386)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2369..2386)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2596..2620)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2617..2820)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2646..2666)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2821..3200
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3367..3383
                     /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(3839..3856)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3839..3855)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3954..3971)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4125..4713)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4205..4224)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4799..4818)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4824..4843)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4885..4976
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4977..5045,5046..5837)
                     /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(5195..5214)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5890..6011
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5901..5920)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5955..5974
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6141..6670,1..70))
                     /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(6224..6244)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cgagggtgcg tacggccctg gggacgtcgt cgcgggtggc gaggcgcacc gtgggcttgt
       61 actcggtcat ggtttagttc ctcaccttgt cgtattatac tatgccgata tactatgccg
      121 atgattaatt gtcaacacgt gctgatcaga tccgaaaatg gatatacaag ctcccgggag
      181 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
      241 ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
      301 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
      361 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
      421 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
      481 gcctggggac tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat
      541 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
      601 tcaaaagaat agaccgagat agggttgagt gttgtgatgc aatttcctca ttttattagg
      661 aaaggacagt gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa
      721 cagatggctg gcaactagaa ggcacagtcg aggctgattt tctctatcta tttattttac
      781 tactcaagac tatcttgcaa agtagaagaa ccctgtctgc agtatgacga ccgtgtatac
      841 ccgattggct taccgggtaa cgcgttgtgc cagtgactac aggccggcct tagtggtgat
      901 ggtggtggtg ttcatgccac tcaatcttct gagcctcgaa aatatcatta agaccgctgc
      961 cgccggggct caggctcagg gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca
     1021 cgctgcagct gaacacgttg ccctgctgcc acctggactt gtccacggtc agcttgctgt
     1081 acaggaagaa gctgccgtcg ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg
     1141 gctggccgtt gctctcccac tccacggcga tatcgctggg gtagaagccc ttcaccagac
     1201 aggtcaggga cacctggttc ttggtcatct cctcccggct ggggggcagg gtgtacacct
     1261 ggggctcccg tggctggccc ttggccttgc tgattgtctt ttcgattggg gcagccaggg
     1321 ccttgttgga gactttgcac ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg
     1381 tcagcacgga caccaccctg taggtgctgt tgtactgctc ctctctgggc ttggtcttgg
     1441 cgttgtgcac ctccacgccg tccacgtacc agttgaactt cacctctggg tcctcgtggg
     1501 acacggccac caccacgcag gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg
     1561 gcttgggggg gaacaggaac acggaaggcc ctcccagcag ttctggagct gggcaggggg
     1621 ggcaggtgtg ggtcttgtcg ccgcccggcg cgccaagaga tggaggcttg gtggggtcca
     1681 gcatctgctc ccagtacatc aggaactctt ccagccacat cttgcagtcg cccagggaga
     1741 tcttctggaa gaacatggtc acgtcccggt tcttctccca cttctcggtc attttcttgg
     1801 cgccagggtg cagagctgtc cacttccggt tgttgctgtc gaacagcagg aacttctggc
     1861 cgttgaacag aaactgccag cttcctcttc cgtggccgtg ggcctcgtgt tcacaagaca
     1921 ttctggcctg cagtgtcagg ggctcgatgg ggatcagatt ttccacctgg atgtccagca
     1981 gctggccctt cagaaaatcc accacgtcgc gcagtgtctc ggtctgttct tcccaggtct
     2041 tggtcacgtt cactttcttg cccaggctgg cgaaggcctt ggccttgtga ttcacacagt
     2101 cgtagtgcag gaagggcctc tcatccacca gtccttgcac ttcgcaccac tgaggctcgg
     2161 gtctgctctt aggggtgatg atgaagtcgt agcacaggca gtgtgtgtca acccatccgg
     2221 cggccgcagc gaacaccagg cagaagatgt agctcagggc aatgatggtc ttcatggtgg
     2281 cgctagcagg cgcgttttcg atagaaccga ggtgcagttg gacctgggag tggacacctg
     2341 tggagagaaa ggcaaagtgg atgttattct atagtgtcac ctaaatcgta tgtgtatgat
     2401 acataaggtt atgtattaat tgtagccgcg ttctaacgaa gccaaggggg tgggcctata
     2461 gactctatag gcggtactta cgtcactctt ggcacgggga atccgcgttc caatgcaccg
     2521 ttcccggccg cggaggctgg atcggtcccg gtgtcttcta tggaggtcaa aacagcgtgg
     2581 atggcgtctc caggcgatct gacggttcac taaacgagct ctgcttatat agacctccca
     2641 ccgtacacgc ctaccgccca tttgcgtcaa tggggcggag ttgttacgac attttggaaa
     2701 gtcccgttga ttttggtgcc aaaacaaact cccattgacg tcaatggggt ggagacttgg
     2761 aaatccccgt gagtcaaacc gctatccacg cccattgatg tactgccaaa accgcatcac
     2821 catggtaata gcgatgacta atacgtagat gtactgccaa gtaggaaagt cccataaggt
     2881 catgtactgg gcataatgcc aggcgggcca tttaccgtca ttgacgtcaa tagggggcgt
     2941 acttggcata tgatacactt gatgtactgc caagtgggca gtttaccgta aatactccac
     3001 ccattgacgt caatggaaag tccctattgg cgttactatg ggaacatacg tcattattga
     3061 cgtcaatggg cgggggtcgt tgggcggtca gccaggcggg ccatttaccg taagttatgt
     3121 aacgcggaac tccatatatg ggctatgaac taatgacccc gtaattgatt actattaata
     3181 actagtcaat aatcaatgcc aacatggcgg tcatattgga catgagccaa tataaatgta
     3241 catattatga tatagataca acgtatgcaa tggccaatag ccaatattga tttatgctat
     3301 ataaccaatg aataatatgg ctaatggcca atattgagtc aacgccactg tgcatgaagc
     3361 tcgtaattgt tatccgctca caattaaggg cgacacaaaa tttattctaa atgataataa
     3421 atactgataa catcttatag tttgtattat attttgtatt atcgttgaca tgtataattt
     3481 tgatatcaaa aactgatttt ccctttatta ttttcgagat ttattttctt aattctcttt
     3541 aacaaactag aaatattgta tatacaaaaa atcataaata atagatgaat agtttaatta
     3601 taggtgttca tcaatcgaaa aagcaacgta tcttatttaa agtgcgttgc ttttttctca
     3661 tttataaggt taaataattc tcatatatca agcaaagtga caggcgccct taaatattct
     3721 gacaaatgct ctttccctaa actcccccca taaaaaaacc cgccgaagcg ggtttttacg
     3781 ttatttgcgg attaacgatt actcgttatc agaaccgccc agggggcccg agcttaagac
     3841 tggccgtcgt tttacaacac agaaagagtt tgtagaaacg caaaaaggcc atccgtcagg
     3901 ggccttctgc ttagtttgat gcctggcagt tccctactct cgccttccgc ttcctcgctc
     3961 actgactcgc tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg
     4021 gtaatacggt tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc
     4081 cagcaaaagg ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc
     4141 ccccctgacg agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga
     4201 ctataaagat accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc
     4261 ctgccgctta ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat
     4321 agctcacgct gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg
     4381 cacgaacccc ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc
     4441 aacccggtaa gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga
     4501 gcgaggtatg taggcggtgc tacagagttc ttgaagtggt gggctaacta cggctacact
     4561 agaagaacag tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt
     4621 ggtagctctt gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag
     4681 cagcagatta cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg
     4741 tctgacgctc agtggaacga cgcgcgcgta actcacgtta agggattttg gtcatgagca
     4801 aggggtgtta tgagccatat tcaggtataa atgggctcgc gataatgttc agaattggtt
     4861 aattggttgt aacactgacc cctatttgtt tatttttcta aatacattca aatatgtatc
     4921 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagaatatga
     4981 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     5041 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     5101 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     5161 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     5221 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     5281 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     5341 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     5401 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     5461 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     5521 tagcgatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     5581 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     5641 cccttccggc tggctggttt attgctgata aatccggagc cggtgagcgt ggttctcgcg
     5701 gtatcatcgc agcgctgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     5761 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     5821 tgattaagca ttggtaatga ccatgattac gccaagctct agctagaggt cgacggtata
     5881 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagtgaaaaa
     5941 aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgtaaccatt ataagctgca
     6001 ataaacaagt tggggtgggc gaagaactcc agcatgagat ccccgcgctg gaggatcatc
     6061 cagccggcgt cccggaaaac gattccgaag cccaaccttt catagaaggc ggcggtggaa
     6121 tcgaaatctc gtagcacgtg tcaggcaccg ggcttgcggg tcatgcacca ggtgcgcggt
     6181 ccttcgggca cctcgacgtc ggcggtgacg gtgaagccga gccgctcgta gaaggggagg
     6241 ttgcggggcg cggatgtctc caggaaggcg ggcaccccgg cgcgctcggc cgcctccact
     6301 ccggggagca cgacggcgct gcccagaccc ttgccctggt ggtcgggcga gacgccgacg
     6361 gtggccagga accacgcggg ctccttgggc cggtgcggcg ccaggaggcc ttccatctgt
     6421 tgctgcgcgg ccagccggga accgctcaac tcggccatgc gcgggccgat ctcggcgaac
     6481 accgcccccg cttcgacgct ctccggcgtg gtccagaccg ccaccgcggc gccgtcgtcc
     6541 gcgacccaca ccttgccgat gtcgagcccg acgcgcgtga ggaagagttc ttgcagctcg
     6601 gtgacccgct cgatgtggcg gtccggatcg acggtgtggc gcgtggcggg gtagtcggcg
     6661 aacgcggcgg
//
