LOCUS       sequence-303039-        8323 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression of cell-surface protein partial 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 8323)
  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 8323)
  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-PAPLN(Trunc3)-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8323
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             5665..5709
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5710..5727
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(5867..5884)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    5873..5984
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6102..6122)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6107..6436
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6287..6422
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6349..6368
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6484..6531
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6550..7149
                     /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(6550..6569)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7046..7066
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7279..7400
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7316..7335)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7370..7389
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7453..8244,8245..8313))
                     /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     8076..8095
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8314..8323,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgcctt gcctcaagag
     2761 gttccatcta tgcaggacgt gcacacccct gccagcaatc cttggatgcc tctgggacct
     2821 caagagagcc ctgcctctga tagtagagga cagtggtggg ccgctcaaga gcatccttct
     2881 gccagaggcg atcacagagg cgaaagggga gatcctagag gcgaccaggg aacacatctg
     2941 tctgctctcg gacctgctcc atctctgcag cagcctccat atcagcagcc actgagatct
     3001 ggcagcggcc ctcacgattg cagacactct ccacacggct gttgtcctga tggccacaca
     3061 gcttctcttg gccctcagtg gcaaggatgt cctggtgctc cttgccagca gagcagatac
     3121 ggctgctgcc ctgatagagt gtcagtggcc gaaggacctc accacgccgg ctgtacaaag
     3181 agctatggcg gagattctac cggcggcatg cctagaagta gagccgtggc ctctaccgtg
     3241 cacaataccc atcagcctca ggctcagcag aacgagccta gcgagtgtag aggcagccag
     3301 ttcggctgct gttacgataa tgtggctaca gccgccggac ctctcggaga gggatgtgtt
     3361 ggacagcctt ctcatgctta ccccgtgcgg tgtctgctgc cttctgctca tggatcttgt
     3421 gccgattggg ccgccagatg gtactttgtg gcttccgtgg gccagtgcaa cagattttgg
     3481 tacggcggat gccacggcaa cgccaacaat tttgcctccg agcaagagtg catgagcagc
     3541 tgtcagggat ctctgcacgg acctagaagg cctcaacctg gcgcctctgg aagatctaca
     3601 cacacagatg gcggaggatc tagccctgcc ggcgaacaag aaccttctca gcatagaaca
     3661 ggcgctgccg tgcaaagaaa gccttggcct tctggcggac tgtggcgaca ggatcaacaa
     3721 cctggacctg gcgaagcccc tcacacacag gcttttggag aatggccttg gggccaagaa
     3781 ctgggatcaa gagcacctgg actcggcgga gatgctggat ctcctgctcc tccatttcac
     3841 agcagcagct acagaatcag cctggctggc gtggaaccat ctctggttca agctgctctg
     3901 ggacagctcg tgcggctgag ctgttctgat gatacagccc ctgaatctca ggccgcctgg
     3961 caaaaagatg gccagccaat cagcagcgac cggcacagac tgcagtttga cggcagcctg
     4021 atcattcacc cactgcaggc tgaggatgcc ggcacataca gctgtggctc taccagacct
     4081 ggccgggatt ctcagaagat ccagctgaga atcatcggcg gcgatatggc cgtgctgtct
     4141 gaagccgaac tcagcagatt ccctcagcct agagatcccg ctcaggattt tggacaagct
     4201 ggcgctgctg gacctctggg agccattcct tctagtcacc ctcagccagc caaccggctg
     4261 agactggacc agaatcaacc cagagtggtg gacgcctctc ctggccagag aatcagaatg
     4321 acatgcagag ccgagggctt cccacctcct gctattgagt ggcagagaga tggacagccc
     4381 gtcagctctc ctagacacca gctgcagcca gatggcagcc tggttattag cagagtggct
     4441 gtggaagatg gcggcttcta cacatgcgtg gccttcaacg gccaggacag agatcagaga
     4501 tgggtgcagc tgagagtgct gggcgagctg acaatttctg gcctgcctcc tacagtgacc
     4561 gtgcctgaag gcgatacagc cagactgctg tgtgtggtgg ccggcgagtc tgtgaacatc
     4621 agatggtcca gaaacggcct gcctgtgcag gccgatggac acagagttca ccagtctcca
     4681 gacggcaccc tgctgatcta caacctgaga gccagagatg agggctccta cacctgtagc
     4741 gcttatcagg gatcacaggc cgtgtccaga tccaccgaag tgaaggttgt gtcccctgct
     4801 cctaccgcac agccacgtga tcctggcaga gattgtgtgg atcagcccga gctggccaac
     4861 tgcgatctga ttctgcaggc ccagctgtgc ggcaacgagt actacagcag cttctgctgc
     4921 gccagctgct ccagatttca gccacacgct cagcccatct ggcaaggcgc gccgggcggc
     4981 gacaagaccc acacctgccc cccctgccca gctccagaac tgctgggagg gccttccgtg
     5041 ttcctgttcc cccccaagcc caaggacacc ctgatgatca gcaggacccc cgaggtgacc
     5101 tgcgtggtgg tggccgtgtc ccacgaggac ccagaggtga agttcaactg gtacgtggac
     5161 ggcgtggagg tgcacaacgc caagaccaag cccagagagg agcagtacaa cagcacctac
     5221 agggtggtgt ccgtgctgac cgtgctgcac caggactggc tgaacggcaa agaatacaag
     5281 tgcaaagtct ccaacaaggc cctggctgcc ccaatcgaaa agacaatcag caaggccaag
     5341 ggccagccac gggagcccca ggtgtacacc ctgcccccca gccgggagga gatgaccaag
     5401 aaccaggtgt ccctgacctg tctggtgaag ggcttctacc ccagcgatat cgccgtggag
     5461 tgggagagca acggccagcc cgagaacaac tacaagacca cccccccagt gctggacagc
     5521 gacggcagct tcttcctgta cagcaagctg accgtggaca agtccaggtg gcagcagggc
     5581 aacgtgttca gctgcagcgt gatgcacgag gccctgcaca accactacac ccagaagtcc
     5641 ctgagcctga gccccggcgg cagcggtctt aatgatattt tcgaggctca gaagattgag
     5701 tggcatgaac accaccacca tcaccactaa ggccggcctg tagtcactgg cacaacgcgt
     5761 tacccggtaa gccaatcggg tatacacggt cgtcatactg cagacagggt tcttctactt
     5821 tgcaagatag tcttgagtag taaaataaat agatagagaa aatcagcctc gactgtgcct
     5881 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     5941 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcacaaca ctcaacccta
     6001 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     6061 atgagctgat ttaacaaaaa tttaacgcga attaattctg tggaatgtgt gtcagttagg
     6121 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     6181 gtcagcaacc aggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     6241 gcatctcaat tagtcagcaa ccatagtccc gcccctaact ccgcccatcc cgcccctaac
     6301 tccgcccagt tccgcccatt ctccgcccca tggctgacta atttttttta tttatgcaga
     6361 ggccgaggcc gcctctgcct ctgagctatt ccagaagtag tgaggaggct tttttggagg
     6421 cctaggcttt tgcaaaaagc tcccgggagc ttgtatatcc attttcggat ctgatcagca
     6481 cgtgttgaca attaatcatc ggcatagtat atcggcatag tataatacga caaggtgagg
     6541 aactaaacca tgaccgagta caagcccacg gtgcgcctcg ccacccgcga cgacgtcccc
     6601 agggccgtac gcaccctcgc cgccgcgttc gccgactacc ccgccacgcg ccacaccgtc
     6661 gatccggacc gccacatcga gcgggtcacc gagctgcaag aactcttcct cacgcgcgtc
     6721 gggctcgaca tcggcaaggt gtgggtcgcg gacgacggcg ccgcggtggc ggtctggacc
     6781 acgccggaga gcgtcgaagc gggggcggtg ttcgccgaga tcggcccgcg catggccgag
     6841 ttgagcggtt cccggctggc cgcgcagcaa cagatggaag gcctcctggc gccgcaccgg
     6901 cccaaggagc ccgcgtggtt cctggccacc gtcggcgtct cgcccgacca ccagggcaag
     6961 ggtctgggca gcgccgtcgt gctccccgga gtggaggcgg ccgagcgcgc cggggtgccc
     7021 gccttcctgg agacatccgc gccccgcaac ctccccttct acgagcggct cggcttcacc
     7081 gtcaccgccg acgtcgaggt gcccgaagga ccgcgcacct ggtgcatgac ccgcaagccc
     7141 ggtgcctgac acgtgctacg agatttcgat tccaccgccg ccttctatga aaggttgggc
     7201 ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga tctcatgctg
     7261 gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa ataaagcaat
     7321 agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg tggtttgtcc
     7381 aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta gagcttggcg
     7441 taatcatggt cattaccaat gcttaatcag tgaggcacct atctcagcga tctgtctatt
     7501 tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatac gggagggctt
     7561 accatctggc cccagcgctg cgatgatacc gcgagaacca cgctcaccgg ctccggattt
     7621 atcagcaata aaccagccag ccggaagggc cgagcgcaga agtggtcctg caactttatc
     7681 cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt cgccagttaa
     7741 tagtttgcgc aacgttgttg ccatcgctac aggcatcgtg gtgtcacgct cgtcgtttgg
     7801 tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat cccccatgtt
     7861 gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gtcagaagta agttggccgc
     7921 agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca tgccatccgt
     7981 aagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat agtgtatgcg
     8041 gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac atagcagaac
     8101 tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga aaactctcaa ggatcttacc
     8161 gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt cagcatcttt
     8221 tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg
     8281 aataagggcg acacggaaat gttgaatact catattcttc ctt
//
