LOCUS       sequence-368606-        6523 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 6523)
  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 6523)
  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-KLRC1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6523
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     107..126
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(145..192)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(240..569)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      254..389
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(308..327)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     554..574
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    692..803
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     792..809
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(949..966)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(967..1011)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2245..2264)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2277..2295)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2278..2295)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2505..2529)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2526..2729)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2555..2575)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2730..3109
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3276..3292
                     /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(3748..3765)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3748..3764)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3863..3880)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4034..4622)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4114..4133)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4708..4727)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4733..4752)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4794..4885
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4886..4954,4955..5746)
                     /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(5104..5123)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5799..5920
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5810..5829)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5864..5883
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6050..6523,1..126))
                     /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(6133..6153)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag
       61 ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc
      121 ggtcatggtt tagttcctca ccttgtcgta ttatactatg ccgatatact atgccgatga
      181 ttaattgtca acacgtgctg atcagatccg aaaatggata tacaagctcc cgggagcttt
      241 ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact tctggaatag ctcagaggca
      301 gaggcggcct cggcctctgc ataaataaaa aaaattagtc agccatgggg cggagaatgg
      361 gcggaactgg gcggagttag gggcgggatg ggcggagtta ggggcgggac tatggttgct
      421 gactaattga gatgcatgct ttgcatactt ctgcctgctg gggagcctgg ggactttcca
      481 cacctggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc tggggagcct
      541 ggggactttc cacaccctaa ctgacacaca ttccacagaa ttaattcgcg ttaaattttt
      601 gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa
      661 aagaatagac cgagataggg ttgagtgttg tgatgcaatt tcctcatttt attaggaaag
      721 gacagtggga gtggcacctt ccagggtcaa ggaaggcacg ggggaggggc aaacaacaga
      781 tggctggcaa ctagaaggca cagtcgaggc tgattttctc tatctattta ttttactact
      841 caagactatc ttgcaaagta gaagaaccct gtctgcagta tgacgaccgt gtatacccga
      901 ttggcttacc gggtaacgcg ttgtgccagt gactacaggc cggccttagt ggtgatggtg
      961 gtggtgttca tgccactcaa tcttctgagc ctcgaaaata tcattaagac cgctgccgcc
     1021 ggggctcagg ctcagggact tctgggtgta gtggttgtgc agggcctcgt gcatcacgct
     1081 gcagctgaac acgttgccct gctgccacct ggacttgtcc acggtcagct tgctgtacag
     1141 gaagaagctg ccgtcgctgt ccagcactgg gggggtggtc ttgtagttgt tctcgggctg
     1201 gccgttgctc tcccactcca cggcgatatc gctggggtag aagcccttca ccagacaggt
     1261 cagggacacc tggttcttgg tcatctcctc ccggctgggg ggcagggtgt acacctgggg
     1321 ctcccgtggc tggcccttgg ccttgctgat tgtcttttcg attggggcag ccagggcctt
     1381 gttggagact ttgcacttgt attctttgcc gttcagccag tcctggtgca gcacggtcag
     1441 cacggacacc accctgtagg tgctgttgta ctgctcctct ctgggcttgg tcttggcgtt
     1501 gtgcacctcc acgccgtcca cgtaccagtt gaacttcacc tctgggtcct cgtgggacac
     1561 ggccaccacc acgcaggtca cctcgggggt cctgctgatc atcagggtgt ccttgggctt
     1621 gggggggaac aggaacacgg aaggccctcc cagcagttct ggagctgggc agggggggca
     1681 ggtgtgggtc ttgtcgccgc ccggcgcgcc aagcttgtgc ttgcagtggt agatgatgct
     1741 gctgccgcac tgggcagact tcagtctgtt cacttgcagc acggcgcaat tcagctcggc
     1801 gttgtcgctg tccttgatct cgtgcttgaa ggccaggccg ttcatggtga cccaaggatg
     1861 gtggctgcta tttctgaaca cgccgatcca gctgctgggg ctgatgatgc tcaggaactt
     1921 catctcttcc tcgttgtcga tgctcagcag ggagctgttc ttgctggtac aggccagcag
     1981 agattcttcc caggtccgcc gctctttgcc gatgtagtag cagctgttgc tgtaggtgat
     2041 ccattcctcg ggacagtgtc cgcagtgtct ggccttctgt gtccgggtgt tcaggctgct
     2101 gttgttgtgt ctctggatca gtgtagaggc ggccgcagcg aacaccaggc agaagatgta
     2161 gctcagggca atgatggtct tcatggtggc gctagcaggc gcgttttcga tagaaccgag
     2221 gtgcagttgg acctgggagt ggacacctgt ggagagaaag gcaaagtgga tgttattcta
     2281 tagtgtcacc taaatcgtat gtgtatgata cataaggtta tgtattaatt gtagccgcgt
     2341 tctaacgaag ccaagggggt gggcctatag actctatagg cggtacttac gtcactcttg
     2401 gcacggggaa tccgcgttcc aatgcaccgt tcccggccgc ggaggctgga tcggtcccgg
     2461 tgtcttctat ggaggtcaaa acagcgtgga tggcgtctcc aggcgatctg acggttcact
     2521 aaacgagctc tgcttatata gacctcccac cgtacacgcc taccgcccat ttgcgtcaat
     2581 ggggcggagt tgttacgaca ttttggaaag tcccgttgat tttggtgcca aaacaaactc
     2641 ccattgacgt caatggggtg gagacttgga aatccccgtg agtcaaaccg ctatccacgc
     2701 ccattgatgt actgccaaaa ccgcatcacc atggtaatag cgatgactaa tacgtagatg
     2761 tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca ggcgggccat
     2821 ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg atgtactgcc
     2881 aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt ccctattggc
     2941 gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt gggcggtcag
     3001 ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg gctatgaact
     3061 aatgaccccg taattgatta ctattaataa ctagtcaata atcaatgcca acatggcggt
     3121 catattggac atgagccaat ataaatgtac atattatgat atagatacaa cgtatgcaat
     3181 ggccaatagc caatattgat ttatgctata taaccaatga ataatatggc taatggccaa
     3241 tattgagtca acgccactgt gcatgaagct cgtaattgtt atccgctcac aattaagggc
     3301 gacacaaaat ttattctaaa tgataataaa tactgataac atcttatagt ttgtattata
     3361 ttttgtatta tcgttgacat gtataatttt gatatcaaaa actgattttc cctttattat
     3421 tttcgagatt tattttctta attctcttta acaaactaga aatattgtat atacaaaaaa
     3481 tcataaataa tagatgaata gtttaattat aggtgttcat caatcgaaaa agcaacgtat
     3541 cttatttaaa gtgcgttgct tttttctcat ttataaggtt aaataattct catatatcaa
     3601 gcaaagtgac aggcgccctt aaatattctg acaaatgctc tttccctaaa ctccccccat
     3661 aaaaaaaccc gccgaagcgg gtttttacgt tatttgcgga ttaacgatta ctcgttatca
     3721 gaaccgccca gggggcccga gcttaagact ggccgtcgtt ttacaacaca gaaagagttt
     3781 gtagaaacgc aaaaaggcca tccgtcaggg gccttctgct tagtttgatg cctggcagtt
     3841 ccctactctc gccttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     3901 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     3961 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     4021 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     4081 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     4141 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     4201 tcccttcggg aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt
     4261 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     4321 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     4381 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     4441 tgaagtggtg ggctaactac ggctacacta gaagaacagt atttggtatc tgcgctctgc
     4501 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     4561 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     4621 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgac gcgcgcgtaa
     4681 ctcacgttaa gggattttgg tcatgagcaa ggggtgttat gagccatatt caggtataaa
     4741 tgggctcgcg ataatgttca gaattggtta attggttgta acactgaccc ctatttgttt
     4801 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     4861 tcaataatat tgaaaaagga agaatatgag tattcaacat ttccgtgtcg cccttattcc
     4921 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     4981 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     5041 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     5101 tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac tcggtcgccg
     5161 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     5221 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     5281 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     5341 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     5401 aaacgacgag cgtgacacca cgatgcctgt agcgatggca acaacgttgc gcaaactatt
     5461 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     5521 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     5581 atccggagcc ggtgagcgtg gttctcgcgg tatcatcgca gcgctggggc cagatggtaa
     5641 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     5701 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaatgac catgattacg
     5761 ccaagctcta gctagaggtc gacggtatac agacatgata agatacattg atgagtttgg
     5821 acaaaccaca actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat
     5881 tgctttattt gtaaccatta taagctgcaa taaacaagtt ggggtgggcg aagaactcca
     5941 gcatgagatc cccgcgctgg aggatcatcc agccggcgtc ccggaaaacg attccgaagc
     6001 ccaacctttc atagaaggcg gcggtggaat cgaaatctcg tagcacgtgt caggcaccgg
     6061 gcttgcgggt catgcaccag gtgcgcggtc cttcgggcac ctcgacgtcg gcggtgacgg
     6121 tgaagccgag ccgctcgtag aaggggaggt tgcggggcgc ggatgtctcc aggaaggcgg
     6181 gcaccccggc gcgctcggcc gcctccactc cggggagcac gacggcgctg cccagaccct
     6241 tgccctggtg gtcgggcgag acgccgacgg tggccaggaa ccacgcgggc tccttgggcc
     6301 ggtgcggcgc caggaggcct tccatctgtt gctgcgcggc cagccgggaa ccgctcaact
     6361 cggccatgcg cgggccgatc tcggcgaaca ccgcccccgc ttcgacgctc tccggcgtgg
     6421 tccagaccgc caccgcggcg ccgtcgtccg cgacccacac cttgccgatg tcgagcccga
     6481 cgcgcgtgag gaagagttct tgcagctcgg tgacccgctc gat
//
