LOCUS       sequence-368737-        6616 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 6616)
  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 6616)
  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-IL23A-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6616
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     103..122
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(141..188)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(236..565)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      250..385
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(304..323)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     550..570
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    688..799
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     788..805
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(945..962)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(963..1007)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2334..2353)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2366..2384)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2367..2384)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2594..2618)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2615..2818)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2644..2664)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2819..3198
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3365..3381
                     /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(3837..3854)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3837..3853)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3952..3969)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4123..4711)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4203..4222)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4797..4816)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4822..4841)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4883..4974
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4975..5043,5044..5835)
                     /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(5193..5212)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5888..6009
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5899..5918)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5953..5972
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6139..6616,1..122))
                     /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(6222..6242)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 cggtccggat cgacggtgtg gcgcgtggcg gggtagtcgg cgaacgcggc ggcgagggtg
       61 cgtacggccc tggggacgtc gtcgcgggtg gcgaggcgca ccgtgggctt gtactcggtc
      121 atggtttagt tcctcacctt gtcgtattat actatgccga tatactatgc cgatgattaa
      181 ttgtcaacac gtgctgatca gatccgaaaa tggatataca agctcccggg agctttttgc
      241 aaaagcctag gcctccaaaa aagcctcctc actacttctg gaatagctca gaggcagagg
      301 cggcctcggc ctctgcataa ataaaaaaaa ttagtcagcc atggggcgga gaatgggcgg
      361 aactgggcgg agttaggggc gggatgggcg gagttagggg cgggactatg gttgctgact
      421 aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac tttccacacc
      481 tggttgctga ctaattgaga tgcatgcttt gcatacttct gcctgctggg gagcctgggg
      541 actttccaca ccctaactga cacacattcc acagaattaa ttcgcgttaa atttttgtta
      601 aatcagctca ttttttaacc aataggccga aatcggcaaa atcccttata aatcaaaaga
      661 atagaccgag atagggttga gtgttgtgat gcaatttcct cattttatta ggaaaggaca
      721 gtgggagtgg caccttccag ggtcaaggaa ggcacggggg aggggcaaac aacagatggc
      781 tggcaactag aaggcacagt cgaggctgat tttctctatc tatttatttt actactcaag
      841 actatcttgc aaagtagaag aaccctgtct gcagtatgac gaccgtgtat acccgattgg
      901 cttaccgggt aacgcgttgt gccagtgact acaggccggc cttagtggtg atggtggtgg
      961 tgttcatgcc actcaatctt ctgagcctcg aaaatatcat taagaccgct gccgccgggg
     1021 ctcaggctca gggacttctg ggtgtagtgg ttgtgcaggg cctcgtgcat cacgctgcag
     1081 ctgaacacgt tgccctgctg ccacctggac ttgtccacgg tcagcttgct gtacaggaag
     1141 aagctgccgt cgctgtccag cactgggggg gtggtcttgt agttgttctc gggctggccg
     1201 ttgctctccc actccacggc gatatcgctg gggtagaagc ccttcaccag acaggtcagg
     1261 gacacctggt tcttggtcat ctcctcccgg ctggggggca gggtgtacac ctggggctcc
     1321 cgtggctggc ccttggcctt gctgattgtc ttttcgattg gggcagccag ggccttgttg
     1381 gagactttgc acttgtattc tttgccgttc agccagtcct ggtgcagcac ggtcagcacg
     1441 gacaccaccc tgtaggtgct gttgtactgc tcctctctgg gcttggtctt ggcgttgtgc
     1501 acctccacgc cgtccacgta ccagttgaac ttcacctctg ggtcctcgtg ggacacggcc
     1561 accaccacgc aggtcacctc gggggtcctg ctgatcatca gggtgtcctt gggcttgggg
     1621 gggaacagga acacggaagg ccctcccagc agttctggag ctgggcaggg ggggcaggtg
     1681 tgggtcttgt cgccgcccgg cgcgccaggg ctcagtgtgg cagcgccatg agcaaaaact
     1741 ctggcagcca cggccacaaa ggcctgcagg gatctcagga tcttgaaccg cagcagcagt
     1801 ctctgccaag gctgagatgg agacagactg gggatctgct gtgtctccca gtggtgtcct
     1861 tcaggctgca gcagttgaga cagtcccagc agagaggcat gcagctgtcc aacaggagaa
     1921 tcaggcagca ggctaggttc gcctgtgaag atgtcgctgc ccagcagctt ctcgtagaag
     1981 atcagtccct ggtgaatccg ctgcagacag aactgggagt tatctctcag gccctgagga
     2041 tcacagccgt cgccacactg gatgtgtggc acgtcgttgg tggtttcttc gtcgccttcc
     2101 tctctcagat ccatgtgtcc cacgagagga tgggcgctcc aagccagtgt acacagcttc
     2161 tggctcagct gctgacactg tgtccaggct ggagaagatc caccaggaac agctctggcg
     2221 gccgcagcga acaccaggca gaagatgtag ctcagggcaa tgatggtctt catggtggcg
     2281 ctagcaggcg cgttttcgat agaaccgagg tgcagttgga cctgggagtg gacacctgtg
     2341 gagagaaagg caaagtggat gttattctat agtgtcacct aaatcgtatg tgtatgatac
     2401 ataaggttat gtattaattg tagccgcgtt ctaacgaagc caagggggtg ggcctataga
     2461 ctctataggc ggtacttacg tcactcttgg cacggggaat ccgcgttcca atgcaccgtt
     2521 cccggccgcg gaggctggat cggtcccggt gtcttctatg gaggtcaaaa cagcgtggat
     2581 ggcgtctcca ggcgatctga cggttcacta aacgagctct gcttatatag acctcccacc
     2641 gtacacgcct accgcccatt tgcgtcaatg gggcggagtt gttacgacat tttggaaagt
     2701 cccgttgatt ttggtgccaa aacaaactcc cattgacgtc aatggggtgg agacttggaa
     2761 atccccgtga gtcaaaccgc tatccacgcc cattgatgta ctgccaaaac cgcatcacca
     2821 tggtaatagc gatgactaat acgtagatgt actgccaagt aggaaagtcc cataaggtca
     2881 tgtactgggc ataatgccag gcgggccatt taccgtcatt gacgtcaata gggggcgtac
     2941 ttggcatatg atacacttga tgtactgcca agtgggcagt ttaccgtaaa tactccaccc
     3001 attgacgtca atggaaagtc cctattggcg ttactatggg aacatacgtc attattgacg
     3061 tcaatgggcg ggggtcgttg ggcggtcagc caggcgggcc atttaccgta agttatgtaa
     3121 cgcggaactc catatatggg ctatgaacta atgaccccgt aattgattac tattaataac
     3181 tagtcaataa tcaatgccaa catggcggtc atattggaca tgagccaata taaatgtaca
     3241 tattatgata tagatacaac gtatgcaatg gccaatagcc aatattgatt tatgctatat
     3301 aaccaatgaa taatatggct aatggccaat attgagtcaa cgccactgtg catgaagctc
     3361 gtaattgtta tccgctcaca attaagggcg acacaaaatt tattctaaat gataataaat
     3421 actgataaca tcttatagtt tgtattatat tttgtattat cgttgacatg tataattttg
     3481 atatcaaaaa ctgattttcc ctttattatt ttcgagattt attttcttaa ttctctttaa
     3541 caaactagaa atattgtata tacaaaaaat cataaataat agatgaatag tttaattata
     3601 ggtgttcatc aatcgaaaaa gcaacgtatc ttatttaaag tgcgttgctt ttttctcatt
     3661 tataaggtta aataattctc atatatcaag caaagtgaca ggcgccctta aatattctga
     3721 caaatgctct ttccctaaac tccccccata aaaaaacccg ccgaagcggg tttttacgtt
     3781 atttgcggat taacgattac tcgttatcag aaccgcccag ggggcccgag cttaagactg
     3841 gccgtcgttt tacaacacag aaagagtttg tagaaacgca aaaaggccat ccgtcagggg
     3901 ccttctgctt agtttgatgc ctggcagttc cctactctcg ccttccgctt cctcgctcac
     3961 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     4021 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     4081 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     4141 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     4201 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     4261 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     4321 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     4381 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     4441 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     4501 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg gctaactacg gctacactag
     4561 aagaacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     4621 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     4681 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     4741 tgacgctcag tggaacgacg cgcgcgtaac tcacgttaag ggattttggt catgagcaag
     4801 gggtgttatg agccatattc aggtataaat gggctcgcga taatgttcag aattggttaa
     4861 ttggttgtaa cactgacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg
     4921 ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa gaatatgagt
     4981 attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt
     5041 gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg
     5101 ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa
     5161 cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt
     5221 gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag
     5281 tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt
     5341 gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga
     5401 ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt
     5461 tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta
     5521 gcgatggcaa caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg
     5581 caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc
     5641 cttccggctg gctggtttat tgctgataaa tccggagccg gtgagcgtgg ttctcgcggt
     5701 atcatcgcag cgctggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg
     5761 gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg
     5821 attaagcatt ggtaatgacc atgattacgc caagctctag ctagaggtcg acggtataca
     5881 gacatgataa gatacattga tgagtttgga caaaccacaa ctagaatgca gtgaaaaaaa
     5941 tgctttattt gtgaaatttg tgatgctatt gctttatttg taaccattat aagctgcaat
     6001 aaacaagttg gggtgggcga agaactccag catgagatcc ccgcgctgga ggatcatcca
     6061 gccggcgtcc cggaaaacga ttccgaagcc caacctttca tagaaggcgg cggtggaatc
     6121 gaaatctcgt agcacgtgtc aggcaccggg cttgcgggtc atgcaccagg tgcgcggtcc
     6181 ttcgggcacc tcgacgtcgg cggtgacggt gaagccgagc cgctcgtaga aggggaggtt
     6241 gcggggcgcg gatgtctcca ggaaggcggg caccccggcg cgctcggccg cctccactcc
     6301 ggggagcacg acggcgctgc ccagaccctt gccctggtgg tcgggcgaga cgccgacggt
     6361 ggccaggaac cacgcgggct ccttgggccg gtgcggcgcc aggaggcctt ccatctgttg
     6421 ctgcgcggcc agccgggaac cgctcaactc ggccatgcgc gggccgatct cggcgaacac
     6481 cgcccccgct tcgacgctct ccggcgtggt ccagaccgcc accgcggcgc cgtcgtccgc
     6541 gacccacacc ttgccgatgt cgagcccgac gcgcgtgagg aagagttctt gcagctcggt
     6601 gacccgctcg atgtgg
//
