LOCUS       sequence-492984-        6955 bp    DNA     circular SYN 18-NOV-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 6955)
  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 6955)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Nov 18, 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-CEACAM6-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6955
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1401..1445
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             1446..1463
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(1603..1620)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1609..1720
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(1838..1858)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1843..2172
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2023..2158
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2085..2104
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        2220..2267
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             2286..2885
                     /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(2286..2305)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     2782..2802
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    3015..3136
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3052..3071)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3106..3125
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(3189..3980,3981..4049))
                     /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     3812..3831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4050..4141)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     4183..4202
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     4208..4227
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      4313..4901
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4802..4821
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5055..5072
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     5170..5187
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     5171..5187
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    5643..5659
                     /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        5826..6205
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        6206..6409
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     6360..6380
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     6406..6430
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        6640..6658
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     6640..6657
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     6671..6690
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
ORIGIN
        1 tcggcacaca gcaagctaca cctggacctg cctacagcgg cagagagaca atctacccta
       61 acgccagcct gctgatccag aacgtgaccc agaacgacac cggcttctac accctgcaag
      121 tgatcaagag cgacctggtc aatgaggaag ccaccggcca gtttcacgtg taccctgagc
      181 tgcctaagcc tagcatcagc agcaacaaca gcaaccccgt ggaagataag gacgccgtgg
      241 cctttacctg cgagcctgag gtgcagaaca ccacctacct ttggtgggtc aacggccagt
      301 ctctgcccgt ttctcctaga ctgcagctga gcaacggcaa catgaccctg acactgctga
      361 gcgtgaagag aaacgatgcc ggcagctacg agtgcgagat ccagaatcct gccagcgcca
      421 atagaagcga ccccgtgaca ctgaatgtgc tgtatggccc tgacggcccc acaatcagcc
      481 ccagcaaagc caattacaga cccggcgaga acctgaacct gagctgtcat gccgccagca
      541 atcctccagc tcagtactcc tggttcatca acggcacctt ccagcagagc acccaagagc
      601 tgttcatccc caacatcacc gtgaacaaca gcggctccta catgtgtcag gcccacaata
      661 gcgccacagg cctgaataga accaccgtga ccatgatcac aggcgcgccg ggcggcgaca
      721 agacccacac ctgccccccc tgcccagctc cagaactgct gggagggcct tccgtgttcc
      781 tgttcccccc caagcccaag gacaccctga tgatcagcag gacccccgag gtgacctgcg
      841 tggtggtggc cgtgtcccac gaggacccag aggtgaagtt caactggtac gtggacggcg
      901 tggaggtgca caacgccaag accaagccca gagaggagca gtacaacagc acctacaggg
      961 tggtgtccgt gctgaccgtg ctgcaccagg actggctgaa cggcaaagaa tacaagtgca
     1021 aagtctccaa caaggccctg gctgccccaa tcgaaaagac aatcagcaag gccaagggcc
     1081 agccacggga gccccaggtg tacaccctgc cccccagccg ggaggagatg accaagaacc
     1141 aggtgtccct gacctgtctg gtgaagggct tctaccccag cgatatcgcc gtggagtggg
     1201 agagcaacgg ccagcccgag aacaactaca agaccacccc cccagtgctg gacagcgacg
     1261 gcagcttctt cctgtacagc aagctgaccg tggacaagtc caggtggcag cagggcaacg
     1321 tgttcagctg cagcgtgatg cacgaggccc tgcacaacca ctacacccag aagtccctga
     1381 gcctgagccc cggcggcagc ggtcttaatg atattttcga ggctcagaag attgagtggc
     1441 atgaacacca ccaccatcac cactaaggcc ggcctgtagt cactggcaca acgcgttacc
     1501 cggtaagcca atcgggtata cacggtcgtc atactgcaga cagggttctt ctactttgca
     1561 agatagtctt gagtagtaaa ataaatagat agagaaaatc agcctcgact gtgccttcta
     1621 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1681 ctcccactgt cctttcctaa taaaatgagg aaattgcatc acaacactca accctatctc
     1741 ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt taaaaaatga
     1801 gctgatttaa caaaaattta acgcgaatta attctgtgga atgtgtgtca gttagggtgt
     1861 ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct caattagtca
     1921 gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat
     1981 ctcaattagt cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg
     2041 cccagttccg cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc
     2101 gaggccgcct ctgcctctga gctattccag aagtagtgag gaggcttttt tggaggccta
     2161 ggcttttgca aaaagctccc gggagcttgt atatccattt tcggatctga tcagcacgtg
     2221 ttgacaatta atcatcggca tagtatatcg gcatagtata atacgacaag gtgaggaact
     2281 aaaccatgac cgagtacaag cccacggtgc gcctcgccac ccgcgacgac gtccccaggg
     2341 ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc cacgcgccac accgtcgatc
     2401 cggaccgcca catcgagcgg gtcaccgagc tgcaagaact cttcctcacg cgcgtcgggc
     2461 tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc ggtggcggtc tggaccacgc
     2521 cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg cccgcgcatg gccgagttga
     2581 gcggttcccg gctggccgcg cagcaacaga tggaaggcct cctggcgccg caccggccca
     2641 aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc cgaccaccag ggcaagggtc
     2701 tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga gcgcgccggg gtgcccgcct
     2761 tcctggagac atccgcgccc cgcaacctcc ccttctacga gcggctcggc ttcaccgtca
     2821 ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg catgacccgc aagcccggtg
     2881 cctgacacgt gctacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     2941 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     3001 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     3061 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     3121 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     3181 catggtcatt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     3241 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     3301 tctggcccca gcgctgcgat gataccgcga gaaccacgct caccggctcc ggatttatca
     3361 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     3421 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     3481 ttgcgcaacg ttgttgccat cgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     3541 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     3601 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     3661 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     3721 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     3781 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     3841 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     3901 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     3961 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     4021 agggcgacac ggaaatgttg aatactcata ttcttccttt ttcaatatta ttgaagcatt
     4081 tatcagggtt attgtctcat gagcggatac atatttgaat gtatttagaa aaataaacaa
     4141 ataggggtca gtgttacaac caattaacca attctgaaca ttatcgcgag cccatttata
     4201 cctgaatatg gctcataaca ccccttgctc atgaccaaaa tcccttaacg tgagttacgc
     4261 gcgcgtcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     4321 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     4381 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     4441 cgcagatacc aaatactgtt cttctagtgt agccgtagtt agcccaccac ttcaagaact
     4501 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     4561 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     4621 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     4681 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     4741 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     4801 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     4861 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     4921 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     4981 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     5041 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga aggcgagagt agggaactgc
     5101 caggcatcaa actaagcaga aggcccctga cggatggcct ttttgcgttt ctacaaactc
     5161 tttctgtgtt gtaaaacgac ggccagtctt aagctcgggc cccctgggcg gttctgataa
     5221 cgagtaatcg ttaatccgca aataacgtaa aaacccgctt cggcgggttt ttttatgggg
     5281 ggagtttagg gaaagagcat ttgtcagaat atttaagggc gcctgtcact ttgcttgata
     5341 tatgagaatt atttaacctt ataaatgaga aaaaagcaac gcactttaaa taagatacgt
     5401 tgctttttcg attgatgaac acctataatt aaactattca tctattattt atgatttttt
     5461 gtatatacaa tatttctagt ttgttaaaga gaattaagaa aataaatctc gaaaataata
     5521 aagggaaaat cagtttttga tatcaaaatt atacatgtca acgataatac aaaatataat
     5581 acaaactata agatgttatc agtatttatt atcatttaga ataaattttg tgtcgccctt
     5641 aattgtgagc ggataacaat tacgagcttc atgcacagtg gcgttgactc aatattggcc
     5701 attagccata ttattcattg gttatatagc ataaatcaat attggctatt ggccattgca
     5761 tacgttgtat ctatatcata atatgtacat ttatattggc tcatgtccaa tatgaccgcc
     5821 atgttggcat tgattattga ctagttatta atagtaatca attacggggt cattagttca
     5881 tagcccatat atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc
     5941 gcccaacgac ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat
     6001 agggactttc cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt
     6061 acatcaagtg tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc
     6121 cgcctggcat tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta
     6181 cgtattagtc atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg
     6241 atagcggttt gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt
     6301 gttttggcac caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac
     6361 gcaaatgggc ggtaggcgtg tacggtggga ggtctatata agcagagctc gtttagtgaa
     6421 ccgtcagatc gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga
     6481 ccgatccagc ctccgcggcc gggaacggtg cattggaacg cggattcccc gtgccaagag
     6541 tgacgtaagt accgcctata gagtctatag gcccaccccc ttggcttcgt tagaacgcgg
     6601 ctacaattaa tacataacct tatgtatcat acacatacga tttaggtgac actatagaat
     6661 aacatccact ttgcctttct ctccacaggt gtccactccc aggtccaact gcacctcggt
     6721 tctatcgaaa acgcgcctgc tagcgccacc atgaagacca tcattgccct gagctacatc
     6781 ttctgcctgg tgttcgctgc ggccgccaag ctgacaatcg agagcacccc ttttaacgtg
     6841 gccgagggca aagaagtgct gctgctggct cacaacctgc ctcagaacag aatcggctac
     6901 agctggtaca agggcgagag agtggacggc aatagcctga tcgtgggcta cgtga
//
