LOCUS       sequence-303079-        8566 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 8566)
  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 8566)
  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-FSTL4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8566
                     /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             5908..5952
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             5953..5970
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6110..6127)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6116..6227
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(6345..6365)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        6350..6679
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      6530..6665
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     6592..6611
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        6727..6774
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             6793..7392
                     /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(6793..6812)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7289..7309
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    7522..7643
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7559..7578)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     7613..7632
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(7696..8487,8488..8556))
                     /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     8319..8338
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8557..8566,1..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 cggccgcctg gatggaccct
     2761 ggaacaagta gaggacctga tgtcggcgtg ggagagtctc aggccgagga acctagaagc
     2821 ttcgaagtga ccagaagaga gggcctgagc agccacaatg agctgctggc ctcttgcggc
     2881 aagaagttct gtagcagagg cagcagatgc gtgctgagca gaaagacagg cgagcctgag
     2941 tgtcagtgcc tggaagcctg tagacctagc tacgtgccag tgtgtggcag cgacggcaga
     3001 ttctacgaga accactgcaa actgcacaga gccgcctgtc tgctgggcaa gagaatcacc
     3061 gtgatccaca gcaaggactg ctttctgaag ggcgatacct gcacaatggc cggctacgcc
     3121 agactgaaaa acgtgctgct ggcactgcag accagactgc aacctctgca agagggcgat
     3181 agtagacagg accccgccag ccagaaaaga ctgctggtgg aaagcctgtt cagagatctg
     3241 gacgccgatg gcaatggcca cctgagttca tctgaactgg cccagcatgt gctgaagaag
     3301 caggacctgg acgaggacct gcttggctgt tctcctggcg acctgctgag attcgacgac
     3361 tacaacagcg acagcagcct gacactgcgc gagttctaca tggcctttca ggtggtgcag
     3421 ctgtctctgg cccctgagga tagagtgtct gtgaccacag tgaccgtggg cctgtctaca
     3481 gtgctgacat gtgctgtgca tggcgatctg cggcctccaa tcatctggaa gagaaacgga
     3541 ctgaccctga acttcctgga cctggaagat atcaacgact tcggcgagga cgacagcctg
     3601 tacatcacca aagtgaccac catccacatg ggcaactaca cctgtcacgc ctctggccac
     3661 gagcagctgt ttcagactca cgtgctgcaa gtgaacgtgc cacctgtgat cagagtgtac
     3721 cccgagagcc aagctcaaga acctggcgtg gcagcctctc tgagatgtca tgccgagggc
     3781 atccccatgc ctagaatcac ctggctgaag aacggcgtgg acgtgtccac acagatgagc
     3841 aaacaactgt ccctgctggc caatggcagc gagctgcaca tcagcagcgt cagatatgag
     3901 gacaccggcg cctacacatg tatcgccaag aatgaagtgg gcgtcgacga ggacatcagc
     3961 tccctgttta tcgaggacag cgccagaaag accctggcca acatcctttg gagagaagag
     4021 ggactgagcg tgggcaacat gttctacgtg ttcagcgacg acggcatcat cgtgattcac
     4081 cccgtggact gcgagatcca gagacacctg aagcctaccg agaagatctt tatgagctac
     4141 gaggaaatct gcccgcagag agagaagaac gccacacagc cttgtcagtg ggtgtcagcc
     4201 gtgaatgtgc ggaaccggta catctatgtg gcccagccag ctctgagtag agtgctggtg
     4261 gtggacattc aggcccagaa ggtgctgcag tctatcggag ttgatcccct gcctgccaag
     4321 ctgtcctacg acaagtctca cgaccaagtg tgggtgctga gctggggcga cgttcacaaa
     4381 agcagaccca gcctgcaagt catcaccgag gcctctacag gacagagcca gcacctgatc
     4441 agaacccctt ttgccggcgt tgacgacttc ttcatccctc ctaccaacct gatcatcaac
     4501 cacatccgct tcggcttcat cttcaacaag agcgaccctg ccgtgcacaa agtggatctg
     4561 gaaaccatga tgcccctgaa aaccatcggc ctgcaccatc acggatgcgt gccacaggct
     4621 atggcccata cacacctcgg cggctacttt ttcatccagt gcagacagga tagccccgcc
     4681 tctgctgcta gacagcttct ggtggatagc gtgaccgata gcgtgctggg acctaatggg
     4741 gatgtgaccg gcacacctca cacaagccct gatggcagat tcatcgtgtc tgccgccgct
     4801 gattctccct ggctgcatgt gcaagagatt acagtgcggg gcgagattca gaccctgtac
     4861 gacctgcaga tcaacagcgg catcagcgac ctggccttcc agagaagctt caccgagagc
     4921 aaccagtaca acatctacgc cgctctgcac accgagcctg acctgctgtt tctggaactg
     4981 agcaccggca aagtgggcat gctgaaaaac ctgaaagagc ctcctgccgg acctgctcaa
     5041 ccttggggag gaacccacag aatcatgaga gatagcggcc tgttcggcca gtacctgctg
     5101 acaccagcca gagagagcct gttcctgatc aacggcagac agaacaccct gagatgcgag
     5161 gtgtccggca tcaaaggcgg cacaacagtc gtgtgggtcg gagaagttgg cgcgccgggc
     5221 ggcgacaaga cccacacctg ccccccctgc ccagctccag aactgctggg agggccttcc
     5281 gtgttcctgt tcccccccaa gcccaaggac accctgatga tcagcaggac ccccgaggtg
     5341 acctgcgtgg tggtggccgt gtcccacgag gacccagagg tgaagttcaa ctggtacgtg
     5401 gacggcgtgg aggtgcacaa cgccaagacc aagcccagag aggagcagta caacagcacc
     5461 tacagggtgg tgtccgtgct gaccgtgctg caccaggact ggctgaacgg caaagaatac
     5521 aagtgcaaag tctccaacaa ggccctggct gccccaatcg aaaagacaat cagcaaggcc
     5581 aagggccagc cacgggagcc ccaggtgtac accctgcccc ccagccggga ggagatgacc
     5641 aagaaccagg tgtccctgac ctgtctggtg aagggcttct accccagcga tatcgccgtg
     5701 gagtgggaga gcaacggcca gcccgagaac aactacaaga ccaccccccc agtgctggac
     5761 agcgacggca gcttcttcct gtacagcaag ctgaccgtgg acaagtccag gtggcagcag
     5821 ggcaacgtgt tcagctgcag cgtgatgcac gaggccctgc acaaccacta cacccagaag
     5881 tccctgagcc tgagccccgg cggcagcggt cttaatgata ttttcgaggc tcagaagatt
     5941 gagtggcatg aacaccacca ccatcaccac taaggccggc ctgtagtcac tggcacaacg
     6001 cgttacccgg taagccaatc gggtatacac ggtcgtcata ctgcagacag ggttcttcta
     6061 ctttgcaaga tagtcttgag tagtaaaata aatagataga gaaaatcagc ctcgactgtg
     6121 ccttctagtt gccagccatc tgttgtttgc ccctcccccg tgccttcctt gaccctggaa
     6181 ggtgccactc ccactgtcct ttcctaataa aatgaggaaa ttgcatcaca acactcaacc
     6241 ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc tattggttaa
     6301 aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     6361 agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca tgcatctcaa
     6421 ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag
     6481 catgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     6541 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     6601 agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag gcttttttgg
     6661 aggcctaggc ttttgcaaaa agctcccggg agcttgtata tccattttcg gatctgatca
     6721 gcacgtgttg acaattaatc atcggcatag tatatcggca tagtataata cgacaaggtg
     6781 aggaactaaa ccatgaccga gtacaagccc acggtgcgcc tcgccacccg cgacgacgtc
     6841 cccagggccg tacgcaccct cgccgccgcg ttcgccgact accccgccac gcgccacacc
     6901 gtcgatccgg accgccacat cgagcgggtc accgagctgc aagaactctt cctcacgcgc
     6961 gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg gcgccgcggt ggcggtctgg
     7021 accacgccgg agagcgtcga agcgggggcg gtgttcgccg agatcggccc gcgcatggcc
     7081 gagttgagcg gttcccggct ggccgcgcag caacagatgg aaggcctcct ggcgccgcac
     7141 cggcccaagg agcccgcgtg gttcctggcc accgtcggcg tctcgcccga ccaccagggc
     7201 aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg cggccgagcg cgccggggtg
     7261 cccgccttcc tggagacatc cgcgccccgc aacctcccct tctacgagcg gctcggcttc
     7321 accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca cctggtgcat gacccgcaag
     7381 cccggtgcct gacacgtgct acgagatttc gattccaccg ccgccttcta tgaaaggttg
     7441 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     7501 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     7561 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     7621 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     7681 gcgtaatcat ggtcattacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     7741 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     7801 cttaccatct ggccccagcg ctgcgatgat accgcgagaa ccacgctcac cggctccgga
     7861 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     7921 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     7981 taatagtttg cgcaacgttg ttgccatcgc tacaggcatc gtggtgtcac gctcgtcgtt
     8041 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     8101 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     8161 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     8221 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     8281 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     8341 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     8401 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     8461 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     8521 gggaataagg gcgacacgga aatgttgaat actcatattc ttcctt
//
