LOCUS       sequence-368739-        6619 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 6619)
  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 6619)
  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-LTA-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6619
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     128..147
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(166..213)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(261..590)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      275..410
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(329..348)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     575..595
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    713..824
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     813..830
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(970..987)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(988..1032)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2362..2381)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2394..2412)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2395..2412)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2622..2646)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2643..2846)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2672..2692)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2847..3226
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3393..3409
                     /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(3865..3882)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3865..3881)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3980..3997)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4151..4739)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4231..4250)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4825..4844)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4850..4869)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4911..5002
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5003..5071,5072..5863)
                     /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(5221..5240)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5916..6037
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5927..5946)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5981..6000
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6167..6619,1..147))
                     /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(6250..6270)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cagctcggtg acccgctcga tgtggcggtc cggatcgacg gtgtggcgcg tggcggggta
       61 gtcggcgaac gcggcggcga gggtgcgtac ggccctgggg acgtcgtcgc gggtggcgag
      121 gcgcaccgtg ggcttgtact cggtcatggt ttagttcctc accttgtcgt attatactat
      181 gccgatatac tatgccgatg attaattgtc aacacgtgct gatcagatcc gaaaatggat
      241 atacaagctc ccgggagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
      301 ttctggaata gctcagaggc agaggcggcc tcggcctctg cataaataaa aaaaattagt
      361 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      421 aggggcggga ctatggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct
      481 ggggagcctg gggactttcc acacctggtt gctgactaat tgagatgcat gctttgcata
      541 cttctgcctg ctggggagcc tggggacttt ccacacccta actgacacac attccacaga
      601 attaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
      661 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gtgatgcaat
      721 ttcctcattt tattaggaaa ggacagtggg agtggcacct tccagggtca aggaaggcac
      781 gggggagggg caaacaacag atggctggca actagaaggc acagtcgagg ctgattttct
      841 ctatctattt attttactac tcaagactat cttgcaaagt agaagaaccc tgtctgcagt
      901 atgacgaccg tgtatacccg attggcttac cgggtaacgc gttgtgccag tgactacagg
      961 ccggccttag tggtgatggt ggtggtgttc atgccactca atcttctgag cctcgaaaat
     1021 atcattaaga ccgctgccgc cggggctcag gctcagggac ttctgggtgt agtggttgtg
     1081 cagggcctcg tgcatcacgc tgcagctgaa cacgttgccc tgctgccacc tggacttgtc
     1141 cacggtcagc ttgctgtaca ggaagaagct gccgtcgctg tccagcactg ggggggtggt
     1201 cttgtagttg ttctcgggct ggccgttgct ctcccactcc acggcgatat cgctggggta
     1261 gaagcccttc accagacagg tcagggacac ctggttcttg gtcatctcct cccggctggg
     1321 gggcagggtg tacacctggg gctcccgtgg ctggcccttg gccttgctga ttgtcttttc
     1381 gattggggca gccagggcct tgttggagac tttgcacttg tattctttgc cgttcagcca
     1441 gtcctggtgc agcacggtca gcacggacac caccctgtag gtgctgttgt actgctcctc
     1501 tctgggcttg gtcttggcgt tgtgcacctc cacgccgtcc acgtaccagt tgaacttcac
     1561 ctctgggtcc tcgtgggaca cggccaccac cacgcaggtc acctcggggg tcctgctgat
     1621 catcagggtg tccttgggct tgggggggaa caggaacacg gaaggccctc ccagcagttc
     1681 tggagctggg cagggggggc aggtgtgggt cttgtcgccg cccggcgcgc caagagcgaa
     1741 ggcgccgaaa aacactgtgc tggggctcag caccaggtga gggattccat ctgtgtgggt
     1801 agacagctga tcgccctggg tcagctgaaa ggcagcgccg tgatacatag agtgcagcca
     1861 gggctcttgc aggccagggt acaccatttt ctggctgctc agcagtggca cgtggaaagg
     1921 gtactgagag ctgaacagct gcacctcgtg agccaggtac agtggagagc ttgtggcctt
     1981 aggagagtag gccttgccgc tgaacaccac ctggctgtac acaaagtaga tgccgctggt
     2041 aggcaccagc aggctgttgt tgctcaggct gaagccatcc tgcagaaagg ctctgtcggt
     2101 attggcccgc cacagcagag aattctgctt gctagggtcg ccgatcagat gagcggctgg
     2161 cttcagtgtg ctgtgggcca gatgcatttt tgggtgctgt ctggctgtct gagcggcaga
     2221 aggtgtaagt ccaacaccag gcaaggcggc cgcagcgaac accaggcaga agatgtagct
     2281 cagggcaatg atggtcttca tggtggcgct agcaggcgcg ttttcgatag aaccgaggtg
     2341 cagttggacc tgggagtgga cacctgtgga gagaaaggca aagtggatgt tattctatag
     2401 tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct
     2461 aacgaagcca agggggtggg cctatagact ctataggcgg tacttacgtc actcttggca
     2521 cggggaatcc gcgttccaat gcaccgttcc cggccgcgga ggctggatcg gtcccggtgt
     2581 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa
     2641 cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2701 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2761 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2821 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2881 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     2941 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     3001 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     3061 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     3121 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     3181 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgccaaca tggcggtcat
     3241 attggacatg agccaatata aatgtacata ttatgatata gatacaacgt atgcaatggc
     3301 caatagccaa tattgattta tgctatataa ccaatgaata atatggctaa tggccaatat
     3361 tgagtcaacg ccactgtgca tgaagctcgt aattgttatc cgctcacaat taagggcgac
     3421 acaaaattta ttctaaatga taataaatac tgataacatc ttatagtttg tattatattt
     3481 tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct ttattatttt
     3541 cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca
     3601 taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt
     3661 atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca
     3721 aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa
     3781 aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa
     3841 ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta
     3901 gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc
     3961 tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     4021 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     4081 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4141 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4201 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4261 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4321 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4381 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4441 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4501 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4561 agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4621 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4681 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4741 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc
     4801 acgttaaggg attttggtca tgagcaaggg gtgttatgag ccatattcag gtataaatgg
     4861 gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt
     4921 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4981 ataatattga aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt
     5041 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     5101 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     5161 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     5221 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     5281 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     5341 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     5401 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     5461 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5521 cgacgagcgt gacaccacga tgcctgtagc gatggcaaca acgttgcgca aactattaac
     5581 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5641 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5701 cggagccggt gagcgtggtt ctcgcggtat catcgcagcg ctggggccag atggtaagcc
     5761 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5821 acagatcgct gagataggtg cctcactgat taagcattgg taatgaccat gattacgcca
     5881 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     5941 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     6001 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     6061 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     6121 acctttcata gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag gcaccgggct
     6181 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
     6241 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga tgtctccagg aaggcgggca
     6301 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
     6361 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
     6421 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
     6481 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
     6541 agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
     6601 gcgtgaggaa gagttcttg
//
