LOCUS       sequence-492983-        8050 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 8050)
  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 8050)
  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-CEACAM5-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..8050
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(66..665)
                     /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(149..169)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     646..665
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(684..731)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(779..1108)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      793..928
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(847..866)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     1093..1113
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    1231..1342
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     1331..1348
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1488..1505)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1506..1550)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(4311..4330)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(4343..4361)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4344..4361)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4571..4595)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(4592..4795)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4621..4641)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        4796..5175
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    5342..5358
                     /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(5814..5831)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(5814..5830)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5929..5946)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6100..6688)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6180..6199)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(6774..6793)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(6799..6818)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        6860..6951
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6952..7020,7021..7812)
                     /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(7170..7189)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    7865..7986
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(7876..7895)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     7930..7949
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaaacgattc cgaagcccaa cctttcatag aaggcggcgg tggaatcgaa atctcgtagc
       61 acgtgtcagg caccgggctt gcgggtcatg caccaggtgc gcggtccttc gggcacctcg
      121 acgtcggcgg tgacggtgaa gccgagccgc tcgtagaagg ggaggttgcg gggcgcggat
      181 gtctccagga aggcgggcac cccggcgcgc tcggccgcct ccactccggg gagcacgacg
      241 gcgctgccca gacccttgcc ctggtggtcg ggcgagacgc cgacggtggc caggaaccac
      301 gcgggctcct tgggccggtg cggcgccagg aggccttcca tctgttgctg cgcggccagc
      361 cgggaaccgc tcaactcggc catgcgcggg ccgatctcgg cgaacaccgc ccccgcttcg
      421 acgctctccg gcgtggtcca gaccgccacc gcggcgccgt cgtccgcgac ccacaccttg
      481 ccgatgtcga gcccgacgcg cgtgaggaag agttcttgca gctcggtgac ccgctcgatg
      541 tggcggtccg gatcgacggt gtggcgcgtg gcggggtagt cggcgaacgc ggcggcgagg
      601 gtgcgtacgg ccctggggac gtcgtcgcgg gtggcgaggc gcaccgtggg cttgtactcg
      661 gtcatggttt agttcctcac cttgtcgtat tatactatgc cgatatacta tgccgatgat
      721 taattgtcaa cacgtgctga tcagatccga aaatggatat acaagctccc gggagctttt
      781 tgcaaaagcc taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag
      841 aggcggcctc ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg
      901 cggaactggg cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg
      961 actaattgag atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     1021 acctggttgc tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg
     1081 gggactttcc acaccctaac tgacacacat tccacagaat taattcgcgt taaatttttg
     1141 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     1201 agaatagacc gagatagggt tgagtgttgt gatgcaattt cctcatttta ttaggaaagg
     1261 acagtgggag tggcaccttc cagggtcaag gaaggcacgg gggaggggca aacaacagat
     1321 ggctggcaac tagaaggcac agtcgaggct gattttctct atctatttat tttactactc
     1381 aagactatct tgcaaagtag aagaaccctg tctgcagtat gacgaccgtg tatacccgat
     1441 tggcttaccg ggtaacgcgt tgtgccagtg actacaggcc ggccttagtg gtgatggtgg
     1501 tggtgttcat gccactcaat cttctgagcc tcgaaaatat cattaagacc gctgccgccg
     1561 gggctcaggc tcagggactt ctgggtgtag tggttgtgca gggcctcgtg catcacgctg
     1621 cagctgaaca cgttgccctg ctgccacctg gacttgtcca cggtcagctt gctgtacagg
     1681 aagaagctgc cgtcgctgtc cagcactggg ggggtggtct tgtagttgtt ctcgggctgg
     1741 ccgttgctct cccactccac ggcgatatcg ctggggtaga agcccttcac cagacaggtc
     1801 agggacacct ggttcttggt catctcctcc cggctggggg gcagggtgta cacctggggc
     1861 tcccgtggct ggcccttggc cttgctgatt gtcttttcga ttggggcagc cagggccttg
     1921 ttggagactt tgcacttgta ttctttgccg ttcagccagt cctggtgcag cacggtcagc
     1981 acggacacca ccctgtaggt gctgttgtac tgctcctctc tgggcttggt cttggcgttg
     2041 tgcacctcca cgccgtccac gtaccagttg aacttcacct ctgggtcctc gtgggacacg
     2101 gccaccacca cgcaggtcac ctcgggggtc ctgctgatca tcagggtgtc cttgggcttg
     2161 ggggggaaca ggaacacgga aggccctccc agcagttctg gagctgggca gggggggcag
     2221 gtgtgggtct tgtcgccgcc cggcgcgcca ccgggagatg tgccggaggc ggacacagtg
     2281 atggacttca cgatggagtt gtttctgccg gtggccagat tggacacgaa gcaggcgtag
     2341 gtgccattgt tgttgggtgt gatcttggcg ataaacagca cctgggtatg ctgctgaggg
     2401 atgccgttga tccgccagga gtactgagga cttgggttgc ttgcgctgtg gcaagacaga
     2461 ttcaggttgg cgccgctcag gtagctgcta tcaggtgggg agatgatggg tgtgtcaggg
     2521 ccatagagca catccagtgt gacagggtcg ctccgattgg cggacacgga attctggatt
     2581 ccgcacacgt aggctctggc gtcgttccgt gtcacattaa acagggtgag tgtccggttg
     2641 ccattagaca gctgcaggcg aggtgacact ggcagtgact ggccgttaac ccaccaaagg
     2701 taagttgtgt tctgagcctc tggttcacag gtgaaggcga cggcatcttt gtcctcaacc
     2761 ggcttggaat tgttagagct gatggaaggt ttaggcagct cggcagacac ggtgattgtt
     2821 ttcactgtgg tccggctgtg gccgctggca gaattgttgg cctggcatgt gtacaggccg
     2881 gagttcttct cggtgatatt gctaatgaag agttcttgtg tgtgctgctg gatattgccg
     2941 tcgatcagcc agctgtactg tgcaggcgga ttggaggcag cgtggcagga cagagacagg
     3001 ttcacgccag gtctgtagta ggtgtagctg gggctgattg tggggtcgtc aggtccatac
     3061 aggacgttca gaatcacggg gtcgctgtgg tccacggaca gcttgttctg aatgccgcac
     3121 tcgtaagggc ccacgtcgtt tctggtcacg ctcagcaggg tcagtgttct gttgtcgttg
     3181 gacagctgga gccggggaga cacggggaga gactgattgt taacccacca caggtatgtg
     3241 gtattctgga tctcaggctc gcatgtcaga gccacggcat cctcgtcctc gacggggtta
     3301 gagttgttgg aggtgatgaa aggctttgga ggctcggcgt acactgtgat ggtggtcacg
     3361 gtggttctgt tcaggccggt atcgctgttg tgggcctgac aggtatagct gccgctgttg
     3421 ttcacggtga tgttggggat gaacagctct tgggtgctct gctggaaggt gccgttcacg
     3481 aaccaactgt actgagctgg aggattgctg gcggcgtgac agctcaggtt caggttctcg
     3541 ccgcttctgt agctggtgtt cagtggagag attgtgggag cgtcggggcc gtacagcaca
     3601 ttcaggatca cgctatcgct ccgcctggca gacactgggt tctgtgtctc gcacttgtag
     3661 ctggcggtgt cattccgggt cacgttgaac agtgtcaggg tccgattgcc gttgctcagc
     3721 tgcagtctgg gagacacagg caggctctgg ttgttgaccc accaaagata ggtggcgtcc
     3781 tgtgtttcgg gctcgcaggt aaaggccacg gcgtccttat cttccacggg cttgctgttg
     3841 ttgctgctga tgctaggctt gggcagctca gggtacactc tgaactggcc ggtggcttcc
     3901 tcgttgacca ggtcgctctt gatcacgtgc agggtgtaga agccggtgtc gttctggatg
     3961 atgttctgga tcagcaggct ggcgttgggg tagatgatct ctctgccgct ataggcaggt
     4021 ccaggtgtag cctgctgggt gccgatcacg tagccgatga tctgtctgtt gccgtccact
     4081 ctctcgccct tgtaccagct gtagccaaac aggtgctgag gcaggttgtg gaccagcagc
     4141 agcacttctt tgccctcggc cacgttgaaa ggggtgctct cgattgtcag cttggcggcc
     4201 gcagcgaaca ccaggcagaa gatgtagctc agggcaatga tggtcttcat ggtggcgcta
     4261 gcaggcgcgt tttcgataga accgaggtgc agttggacct gggagtggac acctgtggag
     4321 agaaaggcaa agtggatgtt attctatagt gtcacctaaa tcgtatgtgt atgatacata
     4381 aggttatgta ttaattgtag ccgcgttcta acgaagccaa gggggtgggc ctatagactc
     4441 tataggcggt acttacgtca ctcttggcac ggggaatccg cgttccaatg caccgttccc
     4501 ggccgcggag gctggatcgg tcccggtgtc ttctatggag gtcaaaacag cgtggatggc
     4561 gtctccaggc gatctgacgg ttcactaaac gagctctgct tatatagacc tcccaccgta
     4621 cacgcctacc gcccatttgc gtcaatgggg cggagttgtt acgacatttt ggaaagtccc
     4681 gttgattttg gtgccaaaac aaactcccat tgacgtcaat ggggtggaga cttggaaatc
     4741 cccgtgagtc aaaccgctat ccacgcccat tgatgtactg ccaaaaccgc atcaccatgg
     4801 taatagcgat gactaatacg tagatgtact gccaagtagg aaagtcccat aaggtcatgt
     4861 actgggcata atgccaggcg ggccatttac cgtcattgac gtcaataggg ggcgtacttg
     4921 gcatatgata cacttgatgt actgccaagt gggcagttta ccgtaaatac tccacccatt
     4981 gacgtcaatg gaaagtccct attggcgtta ctatgggaac atacgtcatt attgacgtca
     5041 atgggcgggg gtcgttgggc ggtcagccag gcgggccatt taccgtaagt tatgtaacgc
     5101 ggaactccat atatgggcta tgaactaatg accccgtaat tgattactat taataactag
     5161 tcaataatca atgccaacat ggcggtcata ttggacatga gccaatataa atgtacatat
     5221 tatgatatag atacaacgta tgcaatggcc aatagccaat attgatttat gctatataac
     5281 caatgaataa tatggctaat ggccaatatt gagtcaacgc cactgtgcat gaagctcgta
     5341 attgttatcc gctcacaatt aagggcgaca caaaatttat tctaaatgat aataaatact
     5401 gataacatct tatagtttgt attatatttt gtattatcgt tgacatgtat aattttgata
     5461 tcaaaaactg attttccctt tattattttc gagatttatt ttcttaattc tctttaacaa
     5521 actagaaata ttgtatatac aaaaaatcat aaataataga tgaatagttt aattataggt
     5581 gttcatcaat cgaaaaagca acgtatctta tttaaagtgc gttgcttttt tctcatttat
     5641 aaggttaaat aattctcata tatcaagcaa agtgacaggc gcccttaaat attctgacaa
     5701 atgctctttc cctaaactcc ccccataaaa aaacccgccg aagcgggttt ttacgttatt
     5761 tgcggattaa cgattactcg ttatcagaac cgcccagggg gcccgagctt aagactggcc
     5821 gtcgttttac aacacagaaa gagtttgtag aaacgcaaaa aggccatccg tcaggggcct
     5881 tctgcttagt ttgatgcctg gcagttccct actctcgcct tccgcttcct cgctcactga
     5941 ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat
     6001 acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca
     6061 aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc
     6121 tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata
     6181 aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc
     6241 gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc
     6301 acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga
     6361 accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc
     6421 ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag
     6481 gtatgtaggc ggtgctacag agttcttgaa gtggtgggct aactacggct acactagaag
     6541 aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag
     6601 ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca
     6661 gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga
     6721 cgctcagtgg aacgacgcgc gcgtaactca cgttaaggga ttttggtcat gagcaagggg
     6781 tgttatgagc catattcagg tataaatggg ctcgcgataa tgttcagaat tggttaattg
     6841 gttgtaacac tgacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc
     6901 atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagaa tatgagtatt
     6961 caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct
     7021 cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt
     7081 tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt
     7141 tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac
     7201 gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac
     7261 tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct
     7321 gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg
     7381 aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg
     7441 gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagcg
     7501 atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa
     7561 caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt
     7621 ccggctggct ggtttattgc tgataaatcc ggagccggtg agcgtggttc tcgcggtatc
     7681 atcgcagcgc tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg
     7741 agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt
     7801 aagcattggt aatgaccatg attacgccaa gctctagcta gaggtcgacg gtatacagac
     7861 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     7921 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     7981 caagttgggg tgggcgaaga actccagcat gagatccccg cgctggagga tcatccagcc
     8041 ggcgtcccgg
//
