LOCUS       sequence-368830-        6478 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 6478)
  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 6478)
  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-MPZ-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6478
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     216..236
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    354..465
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     454..471
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(611..628)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(629..673)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1862..1881)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1894..1912)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1895..1912)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2122..2146)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2143..2346)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2172..2192)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2347..2726
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2893..2909
                     /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(3365..3382)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3365..3381)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3480..3497)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3651..4239)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3731..3750)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4325..4344)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4350..4369)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4411..4502
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4503..4571,4572..5363)
                     /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(4721..4740)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5416..5537
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5427..5446)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5481..5500
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5667..6266)
                     /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(5750..5770)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6247..6266
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6285..6332)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6380..6478,1..231))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6394..6478,1..51)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6448..6467)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
ORIGIN
        1 tcagccatgg ggcggagaat gggcggaact gggcggagtt aggggcggga tgggcggagt
       61 taggggcggg actatggttg ctgactaatt gagatgcatg ctttgcatac ttctgcctgc
      121 tggggagcct ggggactttc cacacctggt tgctgactaa ttgagatgca tgctttgcat
      181 acttctgcct gctggggagc ctggggactt tccacaccct aactgacaca cattccacag
      241 aattaattcg cgttaaattt ttgttaaatc agctcatttt ttaaccaata ggccgaaatc
      301 ggcaaaatcc cttataaatc aaaagaatag accgagatag ggttgagtgt tgtgatgcaa
      361 tttcctcatt ttattaggaa aggacagtgg gagtggcacc ttccagggtc aaggaaggca
      421 cgggggaggg gcaaacaaca gatggctggc aactagaagg cacagtcgag gctgattttc
      481 tctatctatt tattttacta ctcaagacta tcttgcaaag tagaagaacc ctgtctgcag
      541 tatgacgacc gtgtataccc gattggctta ccgggtaacg cgttgtgcca gtgactacag
      601 gccggcctta gtggtgatgg tggtggtgtt catgccactc aatcttctga gcctcgaaaa
      661 tatcattaag accgctgccg ccggggctca ggctcaggga cttctgggtg tagtggttgt
      721 gcagggcctc gtgcatcacg ctgcagctga acacgttgcc ctgctgccac ctggacttgt
      781 ccacggtcag cttgctgtac aggaagaagc tgccgtcgct gtccagcact gggggggtgg
      841 tcttgtagtt gttctcgggc tggccgttgc tctcccactc cacggcgata tcgctggggt
      901 agaagccctt caccagacag gtcagggaca cctggttctt ggtcatctcc tcccggctgg
      961 ggggcagggt gtacacctgg ggctcccgtg gctggccctt ggccttgctg attgtctttt
     1021 cgattggggc agccagggcc ttgttggaga ctttgcactt gtattctttg ccgttcagcc
     1081 agtcctggtg cagcacggtc agcacggaca ccaccctgta ggtgctgttg tactgctcct
     1141 ctctgggctt ggtcttggcg ttgtgcacct ccacgccgtc cacgtaccag ttgaacttca
     1201 cctctgggtc ctcgtgggac acggccacca ccacgcaggt cacctcgggg gtcctgctga
     1261 tcatcagggt gtccttgggc ttggggggga acaggaacac ggaaggccct cccagcagtt
     1321 ctggagctgg gcaggggggg caggtgtggg tcttgtcgcc gcccggcgcg cctctggttg
     1381 gcaccttctc gaacacgtac agggtcactt ggctggtctt gcccacgatg tcaggagggt
     1441 tcttcacgtc gcaggtaaag gtgccgttgt cgctgtagtc caggttgtgg atcacgatag
     1501 atccatcctt ccaccgggga tctccgaccc actggattct ttctttgaag gtgcccactt
     1561 cgtcgatgta aggctggccc ttggcatagt ggaagatgct gatggcatct cttccgcctt
     1621 cgggctgata ccgccaggta aagctgatgt cgtcggacac ccactcgctg ctccaaaagc
     1681 tacagtgcag tgtcactctg ctgcccacgg cgccatgcac ttctctatct gtgtacacca
     1741 cgatggcggc cgcagcgaac accaggcaga agatgtagct cagggcaatg atggtcttca
     1801 tggtggcgct agcaggcgcg ttttcgatag aaccgaggtg cagttggacc tgggagtgga
     1861 cacctgtgga gagaaaggca aagtggatgt tattctatag tgtcacctaa atcgtatgtg
     1921 tatgatacat aaggttatgt attaattgta gccgcgttct aacgaagcca agggggtggg
     1981 cctatagact ctataggcgg tacttacgtc actcttggca cggggaatcc gcgttccaat
     2041 gcaccgttcc cggccgcgga ggctggatcg gtcccggtgt cttctatgga ggtcaaaaca
     2101 gcgtggatgg cgtctccagg cgatctgacg gttcactaaa cgagctctgc ttatatagac
     2161 ctcccaccgt acacgcctac cgcccatttg cgtcaatggg gcggagttgt tacgacattt
     2221 tggaaagtcc cgttgatttt ggtgccaaaa caaactccca ttgacgtcaa tggggtggag
     2281 acttggaaat ccccgtgagt caaaccgcta tccacgccca ttgatgtact gccaaaaccg
     2341 catcaccatg gtaatagcga tgactaatac gtagatgtac tgccaagtag gaaagtccca
     2401 taaggtcatg tactgggcat aatgccaggc gggccattta ccgtcattga cgtcaatagg
     2461 gggcgtactt ggcatatgat acacttgatg tactgccaag tgggcagttt accgtaaata
     2521 ctccacccat tgacgtcaat ggaaagtccc tattggcgtt actatgggaa catacgtcat
     2581 tattgacgtc aatgggcggg ggtcgttggg cggtcagcca ggcgggccat ttaccgtaag
     2641 ttatgtaacg cggaactcca tatatgggct atgaactaat gaccccgtaa ttgattacta
     2701 ttaataacta gtcaataatc aatgccaaca tggcggtcat attggacatg agccaatata
     2761 aatgtacata ttatgatata gatacaacgt atgcaatggc caatagccaa tattgattta
     2821 tgctatataa ccaatgaata atatggctaa tggccaatat tgagtcaacg ccactgtgca
     2881 tgaagctcgt aattgttatc cgctcacaat taagggcgac acaaaattta ttctaaatga
     2941 taataaatac tgataacatc ttatagtttg tattatattt tgtattatcg ttgacatgta
     3001 taattttgat atcaaaaact gattttccct ttattatttt cgagatttat tttcttaatt
     3061 ctctttaaca aactagaaat attgtatata caaaaaatca taaataatag atgaatagtt
     3121 taattatagg tgttcatcaa tcgaaaaagc aacgtatctt atttaaagtg cgttgctttt
     3181 ttctcattta taaggttaaa taattctcat atatcaagca aagtgacagg cgcccttaaa
     3241 tattctgaca aatgctcttt ccctaaactc cccccataaa aaaacccgcc gaagcgggtt
     3301 tttacgttat ttgcggatta acgattactc gttatcagaa ccgcccaggg ggcccgagct
     3361 taagactggc cgtcgtttta caacacagaa agagtttgta gaaacgcaaa aaggccatcc
     3421 gtcaggggcc ttctgcttag tttgatgcct ggcagttccc tactctcgcc ttccgcttcc
     3481 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     3541 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     3601 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     3661 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3721 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3781 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3841 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3901 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3961 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     4021 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtgggc taactacggc
     4081 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     4141 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     4201 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     4261 acggggtctg acgctcagtg gaacgacgcg cgcgtaactc acgttaaggg attttggtca
     4321 tgagcaaggg gtgttatgag ccatattcag gtataaatgg gctcgcgata atgttcagaa
     4381 ttggttaatt ggttgtaaca ctgaccccta tttgtttatt tttctaaata cattcaaata
     4441 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     4501 atatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     4561 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     4621 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     4681 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     4741 cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     4801 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     4861 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     4921 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     4981 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     5041 tgcctgtagc gatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     5101 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     5161 gctcggccct tccggctggc tggtttattg ctgataaatc cggagccggt gagcgtggtt
     5221 ctcgcggtat catcgcagcg ctggggccag atggtaagcc ctcccgtatc gtagttatct
     5281 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     5341 cctcactgat taagcattgg taatgaccat gattacgcca agctctagct agaggtcgac
     5401 ggtatacaga catgataaga tacattgatg agtttggaca aaccacaact agaatgcagt
     5461 gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc tttatttgta accattataa
     5521 gctgcaataa acaagttggg gtgggcgaag aactccagca tgagatcccc gcgctggagg
     5581 atcatccagc cggcgtcccg gaaaacgatt ccgaagccca acctttcata gaaggcggcg
     5641 gtggaatcga aatctcgtag cacgtgtcag gcaccgggct tgcgggtcat gcaccaggtg
     5701 cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga agccgagccg ctcgtagaag
     5761 gggaggttgc ggggcgcgga tgtctccagg aaggcgggca ccccggcgcg ctcggccgcc
     5821 tccactccgg ggagcacgac ggcgctgccc agacccttgc cctggtggtc gggcgagacg
     5881 ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt gcggcgccag gaggccttcc
     5941 atctgttgct gcgcggccag ccgggaaccg ctcaactcgg ccatgcgcgg gccgatctcg
     6001 gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc agaccgccac cgcggcgccg
     6061 tcgtccgcga cccacacctt gccgatgtcg agcccgacgc gcgtgaggaa gagttcttgc
     6121 agctcggtga cccgctcgat gtggcggtcc ggatcgacgg tgtggcgcgt ggcggggtag
     6181 tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga cgtcgtcgcg ggtggcgagg
     6241 cgcaccgtgg gcttgtactc ggtcatggtt tagttcctca ccttgtcgta ttatactatg
     6301 ccgatatact atgccgatga ttaattgtca acacgtgctg atcagatccg aaaatggata
     6361 tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc aaaaaagcct cctcactact
     6421 tctggaatag ctcagaggca gaggcggcct cggcctctgc ataaataaaa aaaattag
//
