LOCUS       sequence-401037-        7288 bp    DNA     circular SYN 15-DEC-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 7288)
  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 7288)
  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-CEACAM1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7288
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        77..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     581..605
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        815..833
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     815..832
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     846..865
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             2864..2908
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2909..2926
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3066..3083)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3072..3183
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3301..3321)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3306..3635
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3486..3621
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3548..3567
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3683..3730
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3749..4348
                     /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(3749..3768)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4245..4265
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4478..4599
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4515..4534)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4569..4588
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4652..5443,5444..5512))
                     /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     5275..5294
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5513..5604)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5646..5665
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5671..5690
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5776..6364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6265..6284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6518..6535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6633..6650
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6634..6650
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7106..7122
                     /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)."
ORIGIN
        1 ggcattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctcgttta gtgaaccgtc
      601 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
      661 ccagcctccg cggccgggaa cggtgcattg gaacgcggat tccccgtgcc aagagtgacg
      721 taagtaccgc ctatagagtc tataggccca cccccttggc ttcgttagaa cgcggctaca
      781 attaatacat aaccttatgt atcatacaca tacgatttag gtgacactat agaataacat
      841 ccactttgcc tttctctcca caggtgtcca ctcccaggtc caactgcacc tcggttctat
      901 cgaaaacgcg cctgctagcg ccaccatgaa gaccatcatt gccctgagct acatcttctg
      961 cctggtgttc gctgcggccg cccaactgac aacagagtcc atgcctttca acgtggccga
     1021 gggcaaagag gtgctgctgc tggttcacaa cctgcctcag cagctgttcg gctacagctg
     1081 gtacaagggc gagagagtgg acggcaacag acagatcgtg ggctacgcca tcggaaccca
     1141 gcaagctaca cctggacctg ccaacagcgg cagagagaca atctacccta acgccagcct
     1201 gctgatccag aacgtgaccc agaacgacac cggcttctac accctgcaag tgatcaagag
     1261 cgacctggtc aacgaggaag ccaccggcca gtttcacgtg taccctgagc tgcccaagcc
     1321 tagcatcagc agcaacaaca gcaaccccgt ggaagataag gacgccgtgg cctttacctg
     1381 cgagcccgag acacaggata ccacctacct gtggtggatc aacaaccaga gcctgccagt
     1441 gtctcccaga ctgcagctga gcaacggcaa tcggacactg acactgctga gcgtgaccag
     1501 aaacgataca ggcccctacg agtgcgagat tcagaacccc gtgtccgcca atagaagcga
     1561 ccccgtgaca ctgaacgtga catacggccc cgacacaccc acaatcagcc ccagcgatac
     1621 ctactacaga cctggcgcca atctgagcct gtcctgctac gccgcttcca atcctcctgc
     1681 tcagtacagc tggctgatca atggcacctt tcagcagagc acccaagagc tgttcatccc
     1741 caacatcacc gtgaacaact ccggcagcta cacatgccac gccaacaata gcgtgaccgg
     1801 ctgcaaccgg accaccgtga aaaccatcat cgtgaccgag ctgagccccg tggtggccaa
     1861 gcctcagatt aaggccagca agaccacagt gaccggcgac aaggatagcg tgaacctgac
     1921 ctgcagcacc aacgataccg gcatcagcat ccggtggttc ttcaagaatc agtccctgcc
     1981 tagctccgag cggatgaagc tgtctcaggg caacaccaca ctgagcatca accctgtgaa
     2041 aagagaggac gccggcacct attggtgcga ggtgttcaac cccatcagca agaaccagag
     2101 cgaccccatc atgctgaatg tgaactacaa cgccctgcct caagagaacg gactgtctcc
     2161 cggtggcgcg ccgggcggcg acaagaccca cacctgcccc ccctgcccag ctccagaact
     2221 gctgggaggg ccttccgtgt tcctgttccc ccccaagccc aaggacaccc tgatgatcag
     2281 caggaccccc gaggtgacct gcgtggtggt ggccgtgtcc cacgaggacc cagaggtgaa
     2341 gttcaactgg tacgtggacg gcgtggaggt gcacaacgcc aagaccaagc ccagagagga
     2401 gcagtacaac agcacctaca gggtggtgtc cgtgctgacc gtgctgcacc aggactggct
     2461 gaacggcaaa gaatacaagt gcaaagtctc caacaaggcc ctggctgccc caatcgaaaa
     2521 gacaatcagc aaggccaagg gccagccacg ggagccccag gtgtacaccc tgccccccag
     2581 ccgggaggag atgaccaaga accaggtgtc cctgacctgt ctggtgaagg gcttctaccc
     2641 cagcgatatc gccgtggagt gggagagcaa cggccagccc gagaacaact acaagaccac
     2701 ccccccagtg ctggacagcg acggcagctt cttcctgtac agcaagctga ccgtggacaa
     2761 gtccaggtgg cagcagggca acgtgttcag ctgcagcgtg atgcacgagg ccctgcacaa
     2821 ccactacacc cagaagtccc tgagcctgag ccccggcggc agcggtctta atgatatttt
     2881 cgaggctcag aagattgagt ggcatgaaca ccaccaccat caccactaag gccggcctgt
     2941 agtcactggc acaacgcgtt acccggtaag ccaatcgggt atacacggtc gtcatactgc
     3001 agacagggtt cttctacttt gcaagatagt cttgagtagt aaaataaata gatagagaaa
     3061 atcagcctcg actgtgcctt ctagttgcca gccatctgtt gtttgcccct cccccgtgcc
     3121 ttccttgacc ctggaaggtg ccactcccac tgtcctttcc taataaaatg aggaaattgc
     3181 atcacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     3241 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt
     3301 ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc
     3361 aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag
     3421 gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc
     3481 cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa
     3541 ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt
     3601 gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca
     3661 ttttcggatc tgatcagcac gtgttgacaa ttaatcatcg gcatagtata tcggcatagt
     3721 ataatacgac aaggtgagga actaaaccat gaccgagtac aagcccacgg tgcgcctcgc
     3781 cacccgcgac gacgtcccca gggccgtacg caccctcgcc gccgcgttcg ccgactaccc
     3841 cgccacgcgc cacaccgtcg atccggaccg ccacatcgag cgggtcaccg agctgcaaga
     3901 actcttcctc acgcgcgtcg ggctcgacat cggcaaggtg tgggtcgcgg acgacggcgc
     3961 cgcggtggcg gtctggacca cgccggagag cgtcgaagcg ggggcggtgt tcgccgagat
     4021 cggcccgcgc atggccgagt tgagcggttc ccggctggcc gcgcagcaac agatggaagg
     4081 cctcctggcg ccgcaccggc ccaaggagcc cgcgtggttc ctggccaccg tcggcgtctc
     4141 gcccgaccac cagggcaagg gtctgggcag cgccgtcgtg ctccccggag tggaggcggc
     4201 cgagcgcgcc ggggtgcccg ccttcctgga gacatccgcg ccccgcaacc tccccttcta
     4261 cgagcggctc ggcttcaccg tcaccgccga cgtcgaggtg cccgaaggac cgcgcacctg
     4321 gtgcatgacc cgcaagcccg gtgcctgaca cgtgctacga gatttcgatt ccaccgccgc
     4381 cttctatgaa aggttgggct tcggaatcgt tttccgggac gccggctgga tgatcctcca
     4441 gcgcggggat ctcatgctgg agttcttcgc ccaccccaac ttgtttattg cagcttataa
     4501 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     4561 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgta taccgtcgac
     4621 ctctagctag agcttggcgt aatcatggtc attaccaatg cttaatcagt gaggcaccta
     4681 tctcagcgat ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa
     4741 ctacgatacg ggagggctta ccatctggcc ccagcgctgc gatgataccg cgagaaccac
     4801 gctcaccggc tccggattta tcagcaataa accagccagc cggaagggcc gagcgcagaa
     4861 gtggtcctgc aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag
     4921 taagtagttc gccagttaat agtttgcgca acgttgttgc catcgctaca ggcatcgtgg
     4981 tgtcacgctc gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag
     5041 ttacatgatc ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg
     5101 tcagaagtaa gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc
     5161 ttactgtcat gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat
     5221 tctgagaata gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata
     5281 ccgcgccaca tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa
     5341 aactctcaag gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca
     5401 actgatcttc agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc
     5461 aaaatgccgc aaaaaaggga ataagggcga cacggaaatg ttgaatactc atattcttcc
     5521 tttttcaata ttattgaagc atttatcagg gttattgtct catgagcgga tacatatttg
     5581 aatgtattta gaaaaataaa caaatagggg tcagtgttac aaccaattaa ccaattctga
     5641 acattatcgc gagcccattt atacctgaat atggctcata acaccccttg ctcatgacca
     5701 aaatccctta acgtgagtta cgcgcgcgtc gttccactga gcgtcagacc ccgtagaaaa
     5761 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     5821 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     5881 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     5941 gttagcccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     6001 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     6061 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     6121 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     6181 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     6241 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     6301 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     6361 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     6421 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     6481 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     6541 ggaaggcgag agtagggaac tgccaggcat caaactaagc agaaggcccc tgacggatgg
     6601 cctttttgcg tttctacaaa ctctttctgt gttgtaaaac gacggccagt cttaagctcg
     6661 ggccccctgg gcggttctga taacgagtaa tcgttaatcc gcaaataacg taaaaacccg
     6721 cttcggcggg tttttttatg gggggagttt agggaaagag catttgtcag aatatttaag
     6781 ggcgcctgtc actttgcttg atatatgaga attatttaac cttataaatg agaaaaaagc
     6841 aacgcacttt aaataagata cgttgctttt tcgattgatg aacacctata attaaactat
     6901 tcatctatta tttatgattt tttgtatata caatatttct agtttgttaa agagaattaa
     6961 gaaaataaat ctcgaaaata ataaagggaa aatcagtttt tgatatcaaa attatacatg
     7021 tcaacgataa tacaaaatat aatacaaact ataagatgtt atcagtattt attatcattt
     7081 agaataaatt ttgtgtcgcc cttaattgtg agcggataac aattacgagc ttcatgcaca
     7141 gtggcgttga ctcaatattg gccattagcc atattattca ttggttatat agcataaatc
     7201 aatattggct attggccatt gcatacgttg tatctatatc ataatatgta catttatatt
     7261 ggctcatgtc caatatgacc gccatgtt
//
