LOCUS       sequence-475004-        7711 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 7711)
  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 7711)
  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-SELE-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7711
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             865..909
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             910..927
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1067..1084)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1073..1184
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1302..1322)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1307..1636
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1487..1622
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1549..1568
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        1684..1731
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             1750..2349
                     /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(1750..1769)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2246..2266
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    2479..2600
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2516..2535)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2570..2589
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(2653..3444,3445..3513))
                     /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     3276..3295
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3514..3605)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     3647..3666
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     3672..3691
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      3777..4365
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4266..4285
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4519..4536
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     4634..4651
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     4635..4651
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5107..5123
                     /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)."
     enhancer        5290..5669
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        5670..5873
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     5824..5844
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     5870..5894
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        6104..6122
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6104..6121
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6135..6154
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 atcaacatga gctgtagcgg cgagcctgtg ttcggcaccg tgtgcaagtt tgcctgtcct
       61 gaaggctgga ccctgaatgg ctctgccgct agaacatgcg gcgccacagg acattggagt
      121 ggcctgctgc ctacatgtga agcccctacc gagagcaaca tccctggcgc gccgggcggc
      181 gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg
      241 ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc
      301 tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac
      361 ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac
      421 agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag
      481 tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag
      541 ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag
      601 aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag
      661 tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc
      721 gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcagggc
      781 aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc
      841 ctgagcctga gccccggcgg cagcggtctt aatgatattt tcgaggctca gaagattgag
      901 tggcatgaac accaccacca tcaccactaa ggccggcctg tagtcactgg cacaacgcgt
      961 tacccggtaa gccaatcggg tatacacggt cgtcatactg cagacagggt tcttctactt
     1021 tgcaagatag tcttgagtag taaaataaat agatagagaa aatcagcctc gactgtgcct
     1081 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     1141 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcacaaca ctcaacccta
     1201 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     1261 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     1321 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     1381 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     1441 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     1501 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     1561 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     1621 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     1681 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     1741 aactaaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     1801 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     1861 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     1921 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     1981 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     2041 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     2101 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     2161 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     2221 gccttcctgg agacatccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     2281 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     2341 ggtgcctgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     2401 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     2461 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     2521 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     2581 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     2641 taatcatggt cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     2701 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     2761 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
     2821 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     2881 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     2941 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     3001 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     3061 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     3121 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     3181 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     3241 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     3301 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     3361 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     3421 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     3481 aataagggcg acacggaaat gttgaatact catattcttc ctttttcaat attattgaag
     3541 catttatcag ggttattgtc tcatgagcgg atacatattt gaatgtattt agaaaaataa
     3601 acaaataggg gtcagtgtta caaccaatta accaattctg aacattatcg cgagcccatt
     3661 tatacctgaa tatggctcat aacacccctt gctcatgacc aaaatccctt aacgtgagtt
     3721 acgcgcgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga
     3781 gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg
     3841 gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc
     3901 agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttagccca ccacttcaag
     3961 aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc
     4021 agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg
     4081 cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac
     4141 accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga
     4201 aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt
     4261 ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag
     4321 cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg
     4381 gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta
     4441 tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc
     4501 agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaaggcga gagtagggaa
     4561 ctgccaggca tcaaactaag cagaaggccc ctgacggatg gcctttttgc gtttctacaa
     4621 actctttctg tgttgtaaaa cgacggccag tcttaagctc gggccccctg ggcggttctg
     4681 ataacgagta atcgttaatc cgcaaataac gtaaaaaccc gcttcggcgg gtttttttat
     4741 ggggggagtt tagggaaaga gcatttgtca gaatatttaa gggcgcctgt cactttgctt
     4801 gatatatgag aattatttaa ccttataaat gagaaaaaag caacgcactt taaataagat
     4861 acgttgcttt ttcgattgat gaacacctat aattaaacta ttcatctatt atttatgatt
     4921 ttttgtatat acaatatttc tagtttgtta aagagaatta agaaaataaa tctcgaaaat
     4981 aataaaggga aaatcagttt ttgatatcaa aattatacat gtcaacgata atacaaaata
     5041 taatacaaac tataagatgt tatcagtatt tattatcatt tagaataaat tttgtgtcgc
     5101 ccttaattgt gagcggataa caattacgag cttcatgcac agtggcgttg actcaatatt
     5161 ggccattagc catattattc attggttata tagcataaat caatattggc tattggccat
     5221 tgcatacgtt gtatctatat cataatatgt acatttatat tggctcatgt ccaatatgac
     5281 cgccatgttg gcattgatta ttgactagtt attaatagta atcaattacg gggtcattag
     5341 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
     5401 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
     5461 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
     5521 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
     5581 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
     5641 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
     5701 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
     5761 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
     5821 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctcgtttag
     5881 tgaaccgtca gatcgcctgg agacgccatc cacgctgttt tgacctccat agaagacacc
     5941 gggaccgatc cagcctccgc ggccgggaac ggtgcattgg aacgcggatt ccccgtgcca
     6001 agagtgacgt aagtaccgcc tatagagtct ataggcccac ccccttggct tcgttagaac
     6061 gcggctacaa ttaatacata accttatgta tcatacacat acgatttagg tgacactata
     6121 gaataacatc cactttgcct ttctctccac aggtgtccac tcccaggtcc aactgcacct
     6181 cggttctatc gaaaacgcgc ctgctagcgc caccatgaag accatcattg ccctgagcta
     6241 catcttctgc ctggtgttcg ctgcggccgc ctggtcttac aataccagca ccgaggccat
     6301 gacctacgat gaggccagcg cctactgcca gcagagatac acacacctgg tggccatcca
     6361 gaacaaagag gaaatcgagt acctgaacag catcctgagc tacagcccca gctactactg
     6421 gatcggcatc cggaaagtga acaacgtgtg ggtctgggtc ggaacccaga agcctctgac
     6481 agaggaagcc aagaattggg cccctggcga gcccaacaac cggcagaaag atgaggactg
     6541 cgtggaaatc tacatcaagc gcgagaagga cgtcggcatg tggaacgacg agcggtgcag
     6601 caagaaaaag ctggccctgt gttacaccgc cgcctgcaca aataccagct gttctggaca
     6661 cggcgagtgt gtggaaacca tcaacaacta cacatgcaag tgcgaccccg gcttcagcgg
     6721 actgaagtgc gagcagatcg tgaactgcac agccctggaa agccctgagc acggctctct
     6781 cgtgtgttct caccctctgg gcaacttcag ctacaacagc agctgcagca tcagctgcga
     6841 cagaggctac ctgcctagca gcatggaaac aatgcagtgc atgagcagcg gcgagtggtc
     6901 cgctcctatt cctgcctgca atgtggtgga atgcgacgcc gtgaccaatc ctgccaacgg
     6961 cttcgtggaa tgctttcaga accccggcag cttcccctgg aacaccacct gtaccttcga
     7021 ctgcgaagag ggcttcgagc tgatgggagc acagagcctg cagtgtacca gcagcggcaa
     7081 ctgggacaac gagaagccta catgcaaggc cgtgacctgc agagccgtta gacagcctca
     7141 gaatggcagc gtgcggtgct ctcattctcc tgccggcgag ttcaccttca agtccagctg
     7201 caacttcacc tgtgaagagg gattcatgct gcaaggcccc gctcaggtgg aatgtaccac
     7261 acaaggacag tggacccagc agatccctgt gtgcgaggcc tttcagtgta ccgctctgag
     7321 caatcccgag cggggctaca tgaactgtct gcctagcgcc agcggctcct tcagatatgg
     7381 aagcagctgc gagttctcct gcgagcaggg ctttgtgctg aagggctcca agagactgca
     7441 gtgcggacct acaggcgagt gggataatga gaagccaacc tgcgaagccg tcagatgtga
     7501 cgccgttcac cagcctccaa agggcctcgt tagatgcgcc cactctccta tcggggagtt
     7561 tacctataag tcctcctgcg cctttagctg cgaggaagga tttgagctgc acggcagcac
     7621 ccagctggaa tgcacatctc aaggccaatg gaccgaggaa gtgcccagct gtcaggtggt
     7681 caagtgtagc tctctggccg tgcctggcaa g
//
