LOCUS       sequence-368818-        6463 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 6463)
  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 6463)
  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-VSTM1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6463
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     102..121
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(140..187)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(235..564)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      249..384
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(303..322)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     549..569
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    687..798
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     787..804
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(944..961)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(962..1006)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2180..2199)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2212..2230)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2213..2230)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2440..2464)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2461..2664)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2490..2510)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2665..3044
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3211..3227
                     /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(3683..3700)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3683..3699)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3798..3815)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3969..4557)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4049..4068)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4643..4662)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4668..4687)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4729..4820
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4821..4889,4890..5681)
                     /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(5039..5058)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5734..5855
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5745..5764)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5799..5818
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(5985..6463,1..121))
                     /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(6068..6088)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 ggtccggatc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg gcgagggtgc
       61 gtacggccct ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg tactcggtca
      121 tggtttagtt cctcaccttg tcgtattata ctatgccgat atactatgcc gatgattaat
      181 tgtcaacacg tgctgatcag atccgaaaat ggatatacaa gctcccggga gctttttgca
      241 aaagcctagg cctccaaaaa agcctcctca ctacttctgg aatagctcag aggcagaggc
      301 ggcctcggcc tctgcataaa taaaaaaaat tagtcagcca tggggcggag aatgggcgga
      361 actgggcgga gttaggggcg ggatgggcgg agttaggggc gggactatgg ttgctgacta
      421 attgagatgc atgctttgca tacttctgcc tgctggggag cctggggact ttccacacct
      481 ggttgctgac taattgagat gcatgctttg catacttctg cctgctgggg agcctgggga
      541 ctttccacac cctaactgac acacattcca cagaattaat tcgcgttaaa tttttgttaa
      601 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      661 tagaccgaga tagggttgag tgttgtgatg caatttcctc attttattag gaaaggacag
      721 tgggagtggc accttccagg gtcaaggaag gcacggggga ggggcaaaca acagatggct
      781 ggcaactaga aggcacagtc gaggctgatt ttctctatct atttatttta ctactcaaga
      841 ctatcttgca aagtagaaga accctgtctg cagtatgacg accgtgtata cccgattggc
      901 ttaccgggta acgcgttgtg ccagtgacta caggccggcc ttagtggtga tggtggtggt
      961 gttcatgcca ctcaatcttc tgagcctcga aaatatcatt aagaccgctg ccgccggggc
     1021 tcaggctcag ggacttctgg gtgtagtggt tgtgcagggc ctcgtgcatc acgctgcagc
     1081 tgaacacgtt gccctgctgc cacctggact tgtccacggt cagcttgctg tacaggaaga
     1141 agctgccgtc gctgtccagc actggggggg tggtcttgta gttgttctcg ggctggccgt
     1201 tgctctccca ctccacggcg atatcgctgg ggtagaagcc cttcaccaga caggtcaggg
     1261 acacctggtt cttggtcatc tcctcccggc tggggggcag ggtgtacacc tggggctccc
     1321 gtggctggcc cttggccttg ctgattgtct tttcgattgg ggcagccagg gccttgttgg
     1381 agactttgca cttgtattct ttgccgttca gccagtcctg gtgcagcacg gtcagcacgg
     1441 acaccaccct gtaggtgctg ttgtactgct cctctctggg cttggtcttg gcgttgtgca
     1501 cctccacgcc gtccacgtac cagttgaact tcacctctgg gtcctcgtgg gacacggcca
     1561 ccaccacgca ggtcacctcg ggggtcctgc tgatcatcag ggtgtccttg ggcttggggg
     1621 ggaacaggaa cacggaaggc cctcccagca gttctggagc tgggcagggg gggcaggtgt
     1681 gggtcttgtc gccgcccggc gcgcctgttc tggtgtcggt cttcatgcta ggggcttcca
     1741 gttcatcgtg cttgtcggtg accaccagct gcagatgttc gctagactcg gaccactcgt
     1801 ggctggctgt tgtcttgtag gcgcagaagt atctgccggc gtccttaggc ttcaggtcgg
     1861 tgaatggaaa ctcggcctca ttctcggcgc tgctctgctc ttgcttgtag ccgctgtcgt
     1921 tcactttccg cagcacgaag gtcacattct ggctgtgggc ctgacacttc agggtcacgt
     1981 tgctctcggc ttccaccaca gaactaggcc aagcgtgcag gctaggctta ggaggcttct
     2041 cgttcttctt ctcgtcctca taggcggccg cagcgaacac caggcagaag atgtagctca
     2101 gggcaatgat ggtcttcatg gtggcgctag caggcgcgtt ttcgatagaa ccgaggtgca
     2161 gttggacctg ggagtggaca cctgtggaga gaaaggcaaa gtggatgtta ttctatagtg
     2221 tcacctaaat cgtatgtgta tgatacataa ggttatgtat taattgtagc cgcgttctaa
     2281 cgaagccaag ggggtgggcc tatagactct ataggcggta cttacgtcac tcttggcacg
     2341 gggaatccgc gttccaatgc accgttcccg gccgcggagg ctggatcggt cccggtgtct
     2401 tctatggagg tcaaaacagc gtggatggcg tctccaggcg atctgacggt tcactaaacg
     2461 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     2521 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     2581 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     2641 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     2701 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     2761 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     2821 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     2881 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     2941 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     3001 ccccgtaatt gattactatt aataactagt caataatcaa tgccaacatg gcggtcatat
     3061 tggacatgag ccaatataaa tgtacatatt atgatataga tacaacgtat gcaatggcca
     3121 atagccaata ttgatttatg ctatataacc aatgaataat atggctaatg gccaatattg
     3181 agtcaacgcc actgtgcatg aagctcgtaa ttgttatccg ctcacaatta agggcgacac
     3241 aaaatttatt ctaaatgata ataaatactg ataacatctt atagtttgta ttatattttg
     3301 tattatcgtt gacatgtata attttgatat caaaaactga ttttcccttt attattttcg
     3361 agatttattt tcttaattct ctttaacaaa ctagaaatat tgtatataca aaaaatcata
     3421 aataatagat gaatagttta attataggtg ttcatcaatc gaaaaagcaa cgtatcttat
     3481 ttaaagtgcg ttgctttttt ctcatttata aggttaaata attctcatat atcaagcaaa
     3541 gtgacaggcg cccttaaata ttctgacaaa tgctctttcc ctaaactccc cccataaaaa
     3601 aacccgccga agcgggtttt tacgttattt gcggattaac gattactcgt tatcagaacc
     3661 gcccaggggg cccgagctta agactggccg tcgttttaca acacagaaag agtttgtaga
     3721 aacgcaaaaa ggccatccgt caggggcctt ctgcttagtt tgatgcctgg cagttcccta
     3781 ctctcgcctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg gctgcggcga
     3841 gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg ggataacgca
     3901 ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa ggccgcgttg
     3961 ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg acgctcaagt
     4021 cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc tggaagctcc
     4081 ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc ctttctccct
     4141 tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc ggtgtaggtc
     4201 gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta
     4261 tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc actggcagca
     4321 gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga gttcttgaag
     4381 tggtgggcta actacggcta cactagaaga acagtatttg gtatctgcgc tctgctgaag
     4441 ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac caccgctggt
     4501 agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg atctcaagaa
     4561 gatcctttga tcttttctac ggggtctgac gctcagtgga acgacgcgcg cgtaactcac
     4621 gttaagggat tttggtcatg agcaaggggt gttatgagcc atattcaggt ataaatgggc
     4681 tcgcgataat gttcagaatt ggttaattgg ttgtaacact gacccctatt tgtttatttt
     4741 tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat
     4801 aatattgaaa aaggaagaat atgagtattc aacatttccg tgtcgccctt attccctttt
     4861 ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg
     4921 ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga
     4981 tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc
     5041 tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac
     5101 actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg
     5161 gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca
     5221 acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg
     5281 gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg
     5341 acgagcgtga caccacgatg cctgtagcga tggcaacaac gttgcgcaaa ctattaactg
     5401 gcgaactact tactctagct tcccggcaac aattaataga ctggatggag gcggataaag
     5461 ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatccg
     5521 gagccggtga gcgtggttct cgcggtatca tcgcagcgct ggggccagat ggtaagccct
     5581 cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac
     5641 agatcgctga gataggtgcc tcactgatta agcattggta atgaccatga ttacgccaag
     5701 ctctagctag aggtcgacgg tatacagaca tgataagata cattgatgag tttggacaaa
     5761 ccacaactag aatgcagtga aaaaaatgct ttatttgtga aatttgtgat gctattgctt
     5821 tatttgtaac cattataagc tgcaataaac aagttggggt gggcgaagaa ctccagcatg
     5881 agatccccgc gctggaggat catccagccg gcgtcccgga aaacgattcc gaagcccaac
     5941 ctttcataga aggcggcggt ggaatcgaaa tctcgtagca cgtgtcaggc accgggcttg
     6001 cgggtcatgc accaggtgcg cggtccttcg ggcacctcga cgtcggcggt gacggtgaag
     6061 ccgagccgct cgtagaaggg gaggttgcgg ggcgcggatg tctccaggaa ggcgggcacc
     6121 ccggcgcgct cggccgcctc cactccgggg agcacgacgg cgctgcccag acccttgccc
     6181 tggtggtcgg gcgagacgcc gacggtggcc aggaaccacg cgggctcctt gggccggtgc
     6241 ggcgccagga ggccttccat ctgttgctgc gcggccagcc gggaaccgct caactcggcc
     6301 atgcgcgggc cgatctcggc gaacaccgcc cccgcttcga cgctctccgg cgtggtccag
     6361 accgccaccg cggcgccgtc gtccgcgacc cacaccttgc cgatgtcgag cccgacgcgc
     6421 gtgaggaaga gttcttgcag ctcggtgacc cgctcgatgt ggc
//
