LOCUS       sequence-368827-        6469 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 6469)
  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 6469)
  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-CEACAM3-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6469
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     196..215
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(234..281)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(329..658)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      343..478
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(397..416)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     643..663
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    781..892
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     881..898
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(1038..1055)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(1056..1100)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(2280..2299)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(2312..2330)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2313..2330)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2540..2564)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2561..2764)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2590..2610)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2765..3144
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    3311..3327
                     /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(3783..3800)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3783..3799)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3898..3915)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4069..4657)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4149..4168)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4743..4762)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4768..4787)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4829..4920
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4921..4989,4990..5781)
                     /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(5139..5158)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5834..5955
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5845..5864)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5899..5918
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(join(6085..6469,1..215))
                     /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(6168..6188)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
ORIGIN
        1 gcggcgccgt cgtccgcgac ccacaccttg ccgatgtcga gcccgacgcg cgtgaggaag
       61 agttcttgca gctcggtgac ccgctcgatg tggcggtccg gatcgacggt gtggcgcgtg
      121 gcggggtagt cggcgaacgc ggcggcgagg gtgcgtacgg ccctggggac gtcgtcgcgg
      181 gtggcgaggc gcaccgtggg cttgtactcg gtcatggttt agttcctcac cttgtcgtat
      241 tatactatgc cgatatacta tgccgatgat taattgtcaa cacgtgctga tcagatccga
      301 aaatggatat acaagctccc gggagctttt tgcaaaagcc taggcctcca aaaaagcctc
      361 ctcactactt ctggaatagc tcagaggcag aggcggcctc ggcctctgca taaataaaaa
      421 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
      481 gcggagttag gggcgggact atggttgctg actaattgag atgcatgctt tgcatacttc
      541 tgcctgctgg ggagcctggg gactttccac acctggttgc tgactaattg agatgcatgc
      601 tttgcatact tctgcctgct ggggagcctg gggactttcc acaccctaac tgacacacat
      661 tccacagaat taattcgcgt taaatttttg ttaaatcagc tcatttttta accaataggc
      721 cgaaatcggc aaaatccctt ataaatcaaa agaatagacc gagatagggt tgagtgttgt
      781 gatgcaattt cctcatttta ttaggaaagg acagtgggag tggcaccttc cagggtcaag
      841 gaaggcacgg gggaggggca aacaacagat ggctggcaac tagaaggcac agtcgaggct
      901 gattttctct atctatttat tttactactc aagactatct tgcaaagtag aagaaccctg
      961 tctgcagtat gacgaccgtg tatacccgat tggcttaccg ggtaacgcgt tgtgccagtg
     1021 actacaggcc ggccttagtg gtgatggtgg tggtgttcat gccactcaat cttctgagcc
     1081 tcgaaaatat cattaagacc gctgccgccg gggctcaggc tcagggactt ctgggtgtag
     1141 tggttgtgca gggcctcgtg catcacgctg cagctgaaca cgttgccctg ctgccacctg
     1201 gacttgtcca cggtcagctt gctgtacagg aagaagctgc cgtcgctgtc cagcactggg
     1261 ggggtggtct tgtagttgtt ctcgggctgg ccgttgctct cccactccac ggcgatatcg
     1321 ctggggtaga agcccttcac cagacaggtc agggacacct ggttcttggt catctcctcc
     1381 cggctggggg gcagggtgta cacctggggc tcccgtggct ggcccttggc cttgctgatt
     1441 gtcttttcga ttggggcagc cagggccttg ttggagactt tgcacttgta ttctttgccg
     1501 ttcagccagt cctggtgcag cacggtcagc acggacacca ccctgtaggt gctgttgtac
     1561 tgctcctctc tgggcttggt cttggcgttg tgcacctcca cgccgtccac gtaccagttg
     1621 aacttcacct ctgggtcctc gtgggacacg gccaccacca cgcaggtcac ctcgggggtc
     1681 ctgctgatca tcagggtgtc cttgggcttg ggggggaaca ggaacacgga aggccctccc
     1741 agcagttctg gagctgggca gggggggcag gtgtgggtct tgtcgccgcc cggcgcgcca
     1801 ccggcaacag ctcccacagg cagtccaggg gcattttctt ggtacacgtg gaactggccg
     1861 gtggcttcct cattgaccag gtcgctcttg atcacttgca gggtgtagaa gccgatgtcg
     1921 ttctgggtca cgttctggat cagcaggctg gcgttggtat agattgtctc tctgccgctg
     1981 taagcggcgc caggtgtagc ttgttgtgtg ccgatcacgt agcccacgat caggctattg
     2041 ccgtccactc tctcgccctt ataccagctg tagccgaaca ggtgctgagg caggttgtga
     2101 accagcagca gcacctcttt tccctcggcc acgctcagag gcatagactc gattgtcagc
     2161 ttggcggccg cagcgaacac caggcagaag atgtagctca gggcaatgat ggtcttcatg
     2221 gtggcgctag caggcgcgtt ttcgatagaa ccgaggtgca gttggacctg ggagtggaca
     2281 cctgtggaga gaaaggcaaa gtggatgtta ttctatagtg tcacctaaat cgtatgtgta
     2341 tgatacataa ggttatgtat taattgtagc cgcgttctaa cgaagccaag ggggtgggcc
     2401 tatagactct ataggcggta cttacgtcac tcttggcacg gggaatccgc gttccaatgc
     2461 accgttcccg gccgcggagg ctggatcggt cccggtgtct tctatggagg tcaaaacagc
     2521 gtggatggcg tctccaggcg atctgacggt tcactaaacg agctctgctt atatagacct
     2581 cccaccgtac acgcctaccg cccatttgcg tcaatggggc ggagttgtta cgacattttg
     2641 gaaagtcccg ttgattttgg tgccaaaaca aactcccatt gacgtcaatg gggtggagac
     2701 ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt gatgtactgc caaaaccgca
     2761 tcaccatggt aatagcgatg actaatacgt agatgtactg ccaagtagga aagtcccata
     2821 aggtcatgta ctgggcataa tgccaggcgg gccatttacc gtcattgacg tcaatagggg
     2881 gcgtacttgg catatgatac acttgatgta ctgccaagtg ggcagtttac cgtaaatact
     2941 ccacccattg acgtcaatgg aaagtcccta ttggcgttac tatgggaaca tacgtcatta
     3001 ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg cgggccattt accgtaagtt
     3061 atgtaacgcg gaactccata tatgggctat gaactaatga ccccgtaatt gattactatt
     3121 aataactagt caataatcaa tgccaacatg gcggtcatat tggacatgag ccaatataaa
     3181 tgtacatatt atgatataga tacaacgtat gcaatggcca atagccaata ttgatttatg
     3241 ctatataacc aatgaataat atggctaatg gccaatattg agtcaacgcc actgtgcatg
     3301 aagctcgtaa ttgttatccg ctcacaatta agggcgacac aaaatttatt ctaaatgata
     3361 ataaatactg ataacatctt atagtttgta ttatattttg tattatcgtt gacatgtata
     3421 attttgatat caaaaactga ttttcccttt attattttcg agatttattt tcttaattct
     3481 ctttaacaaa ctagaaatat tgtatataca aaaaatcata aataatagat gaatagttta
     3541 attataggtg ttcatcaatc gaaaaagcaa cgtatcttat ttaaagtgcg ttgctttttt
     3601 ctcatttata aggttaaata attctcatat atcaagcaaa gtgacaggcg cccttaaata
     3661 ttctgacaaa tgctctttcc ctaaactccc cccataaaaa aacccgccga agcgggtttt
     3721 tacgttattt gcggattaac gattactcgt tatcagaacc gcccaggggg cccgagctta
     3781 agactggccg tcgttttaca acacagaaag agtttgtaga aacgcaaaaa ggccatccgt
     3841 caggggcctt ctgcttagtt tgatgcctgg cagttcccta ctctcgcctt ccgcttcctc
     3901 gctcactgac tcgctgcgct cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa
     3961 ggcggtaata cggttatcca cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa
     4021 aggccagcaa aaggccagga accgtaaaaa ggccgcgttg ctggcgtttt tccataggct
     4081 ccgcccccct gacgagcatc acaaaaatcg acgctcaagt cagaggtggc gaaacccgac
     4141 aggactataa agataccagg cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc
     4201 gaccctgccg cttaccggat acctgtccgc ctttctccct tcgggaagcg tggcgctttc
     4261 tcatagctca cgctgtaggt atctcagttc ggtgtaggtc gttcgctcca agctgggctg
     4321 tgtgcacgaa ccccccgttc agcccgaccg ctgcgcctta tccggtaact atcgtcttga
     4381 gtccaacccg gtaagacacg acttatcgcc actggcagca gccactggta acaggattag
     4441 cagagcgagg tatgtaggcg gtgctacaga gttcttgaag tggtgggcta actacggcta
     4501 cactagaaga acagtatttg gtatctgcgc tctgctgaag ccagttacct tcggaaaaag
     4561 agttggtagc tcttgatccg gcaaacaaac caccgctggt agcggtggtt tttttgtttg
     4621 caagcagcag attacgcgca gaaaaaaagg atctcaagaa gatcctttga tcttttctac
     4681 ggggtctgac gctcagtgga acgacgcgcg cgtaactcac gttaagggat tttggtcatg
     4741 agcaaggggt gttatgagcc atattcaggt ataaatgggc tcgcgataat gttcagaatt
     4801 ggttaattgg ttgtaacact gacccctatt tgtttatttt tctaaataca ttcaaatatg
     4861 tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa aaggaagaat
     4921 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
     4981 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
     5041 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
     5101 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
     5161 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
     5221 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
     5281 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
     5341 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
     5401 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
     5461 cctgtagcga tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
     5521 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
     5581 tcggcccttc cggctggctg gtttattgct gataaatccg gagccggtga gcgtggttct
     5641 cgcggtatca tcgcagcgct ggggccagat ggtaagccct cccgtatcgt agttatctac
     5701 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
     5761 tcactgatta agcattggta atgaccatga ttacgccaag ctctagctag aggtcgacgg
     5821 tatacagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     5881 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     5941 tgcaataaac aagttggggt gggcgaagaa ctccagcatg agatccccgc gctggaggat
     6001 catccagccg gcgtcccgga aaacgattcc gaagcccaac ctttcataga aggcggcggt
     6061 ggaatcgaaa tctcgtagca cgtgtcaggc accgggcttg cgggtcatgc accaggtgcg
     6121 cggtccttcg ggcacctcga cgtcggcggt gacggtgaag ccgagccgct cgtagaaggg
     6181 gaggttgcgg ggcgcggatg tctccaggaa ggcgggcacc ccggcgcgct cggccgcctc
     6241 cactccgggg agcacgacgg cgctgcccag acccttgccc tggtggtcgg gcgagacgcc
     6301 gacggtggcc aggaaccacg cgggctcctt gggccggtgc ggcgccagga ggccttccat
     6361 ctgttgctgc gcggccagcc gggaaccgct caactcggcc atgcgcgggc cgatctcggc
     6421 gaacaccgcc cccgcttcga cgctctccgg cgtggtccag accgccacc
//
