LOCUS       sequence-368821-        6466 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 6466)
  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 6466)
  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-CEACAM4-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..6466
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2..21)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     248..268
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     polyA_signal    386..497
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     486..503
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     CDS             complement(643..660)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             complement(661..705)
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     primer_bind     complement(1882..1901)
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     promoter        complement(1914..1932)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1915..1932)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2142..2166)
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        complement(2163..2366)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2192..2212)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2367..2746
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     protein_bind    2913..2929
                     /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(3385..3402)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(3385..3401)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3500..3517)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3671..4259)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3751..3770)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     complement(4345..4364)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     primer_bind     complement(4370..4389)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     promoter        4431..4522
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4523..4591,4592..5383)
                     /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(4741..4760)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     polyA_signal    5436..5557
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5447..5466)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     5501..5520
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     CDS             complement(5687..6286)
                     /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(5770..5790)
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     primer_bind     6267..6286
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     promoter        complement(6305..6352)
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     promoter        complement(join(6400..6466,1..263))
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      join(6414..6466,1..83)
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga atgggcggaa
       61 ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt tgctgactaa
      121 ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt tccacacctg
      181 gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga gcctggggac
      241 tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat ttttgttaaa
      301 tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat
      361 agaccgagat agggttgagt gttgtgatgc aatttcctca ttttattagg aaaggacagt
      421 gggagtggca ccttccaggg tcaaggaagg cacgggggag gggcaaacaa cagatggctg
      481 gcaactagaa ggcacagtcg aggctgattt tctctatcta tttattttac tactcaagac
      541 tatcttgcaa agtagaagaa ccctgtctgc agtatgacga ccgtgtatac ccgattggct
      601 taccgggtaa cgcgttgtgc cagtgactac aggccggcct tagtggtgat ggtggtggtg
      661 ttcatgccac tcaatcttct gagcctcgaa aatatcatta agaccgctgc cgccggggct
      721 caggctcagg gacttctggg tgtagtggtt gtgcagggcc tcgtgcatca cgctgcagct
      781 gaacacgttg ccctgctgcc acctggactt gtccacggtc agcttgctgt acaggaagaa
      841 gctgccgtcg ctgtccagca ctgggggggt ggtcttgtag ttgttctcgg gctggccgtt
      901 gctctcccac tccacggcga tatcgctggg gtagaagccc ttcaccagac aggtcaggga
      961 cacctggttc ttggtcatct cctcccggct ggggggcagg gtgtacacct ggggctcccg
     1021 tggctggccc ttggccttgc tgattgtctt ttcgattggg gcagccaggg ccttgttgga
     1081 gactttgcac ttgtattctt tgccgttcag ccagtcctgg tgcagcacgg tcagcacgga
     1141 caccaccctg taggtgctgt tgtactgctc ctctctgggc ttggtcttgg cgttgtgcac
     1201 ctccacgccg tccacgtacc agttgaactt cacctctggg tcctcgtggg acacggccac
     1261 caccacgcag gtcacctcgg gggtcctgct gatcatcagg gtgtccttgg gcttgggggg
     1321 gaacaggaac acggaaggcc ctcccagcag ttctggagct gggcaggggg ggcaggtgtg
     1381 ggtcttgtcg ccgcccggcg cgccaccggc cacagctcca acaggcagtc caggcacatt
     1441 gttctggtgc acatgcagct gtcctgtggc ctgatcagag tcgtagctgg cgttgatggt
     1501 tctcagggtg tagctgccgg catcttccag tgtgatgttc tggaacagca ggctgccgtt
     1561 agggtacact gtttctctgc cgctgtaggc agcgccggga atattggcct ggatgtcggt
     1621 gatgtagccg gcgatcagag gagatccctc ggctgtcttg cccttgtgcc agtagtaggc
     1681 ctggattgtc tcgctgatgt tgcaggccag cagcagcaca tcttttccct cggcggcaga
     1741 agatggcagg gcctcaattg taaaggcggc cgcagcgaac accaggcaga agatgtagct
     1801 cagggcaatg atggtcttca tggtggcgct agcaggcgcg ttttcgatag aaccgaggtg
     1861 cagttggacc tgggagtgga cacctgtgga gagaaaggca aagtggatgt tattctatag
     1921 tgtcacctaa atcgtatgtg tatgatacat aaggttatgt attaattgta gccgcgttct
     1981 aacgaagcca agggggtggg cctatagact ctataggcgg tacttacgtc actcttggca
     2041 cggggaatcc gcgttccaat gcaccgttcc cggccgcgga ggctggatcg gtcccggtgt
     2101 cttctatgga ggtcaaaaca gcgtggatgg cgtctccagg cgatctgacg gttcactaaa
     2161 cgagctctgc ttatatagac ctcccaccgt acacgcctac cgcccatttg cgtcaatggg
     2221 gcggagttgt tacgacattt tggaaagtcc cgttgatttt ggtgccaaaa caaactccca
     2281 ttgacgtcaa tggggtggag acttggaaat ccccgtgagt caaaccgcta tccacgccca
     2341 ttgatgtact gccaaaaccg catcaccatg gtaatagcga tgactaatac gtagatgtac
     2401 tgccaagtag gaaagtccca taaggtcatg tactgggcat aatgccaggc gggccattta
     2461 ccgtcattga cgtcaatagg gggcgtactt ggcatatgat acacttgatg tactgccaag
     2521 tgggcagttt accgtaaata ctccacccat tgacgtcaat ggaaagtccc tattggcgtt
     2581 actatgggaa catacgtcat tattgacgtc aatgggcggg ggtcgttggg cggtcagcca
     2641 ggcgggccat ttaccgtaag ttatgtaacg cggaactcca tatatgggct atgaactaat
     2701 gaccccgtaa ttgattacta ttaataacta gtcaataatc aatgccaaca tggcggtcat
     2761 attggacatg agccaatata aatgtacata ttatgatata gatacaacgt atgcaatggc
     2821 caatagccaa tattgattta tgctatataa ccaatgaata atatggctaa tggccaatat
     2881 tgagtcaacg ccactgtgca tgaagctcgt aattgttatc cgctcacaat taagggcgac
     2941 acaaaattta ttctaaatga taataaatac tgataacatc ttatagtttg tattatattt
     3001 tgtattatcg ttgacatgta taattttgat atcaaaaact gattttccct ttattatttt
     3061 cgagatttat tttcttaatt ctctttaaca aactagaaat attgtatata caaaaaatca
     3121 taaataatag atgaatagtt taattatagg tgttcatcaa tcgaaaaagc aacgtatctt
     3181 atttaaagtg cgttgctttt ttctcattta taaggttaaa taattctcat atatcaagca
     3241 aagtgacagg cgcccttaaa tattctgaca aatgctcttt ccctaaactc cccccataaa
     3301 aaaacccgcc gaagcgggtt tttacgttat ttgcggatta acgattactc gttatcagaa
     3361 ccgcccaggg ggcccgagct taagactggc cgtcgtttta caacacagaa agagtttgta
     3421 gaaacgcaaa aaggccatcc gtcaggggcc ttctgcttag tttgatgcct ggcagttccc
     3481 tactctcgcc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3541 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3601 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     3661 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3721 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3781 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3841 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3901 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3961 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4021 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4081 agtggtgggc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4141 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4201 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4261 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgacgcg cgcgtaactc
     4321 acgttaaggg attttggtca tgagcaaggg gtgttatgag ccatattcag gtataaatgg
     4381 gctcgcgata atgttcagaa ttggttaatt ggttgtaaca ctgaccccta tttgtttatt
     4441 tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca
     4501 ataatattga aaaaggaaga atatgagtat tcaacatttc cgtgtcgccc ttattccctt
     4561 ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga
     4621 tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa
     4681 gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct
     4741 gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat
     4801 acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga
     4861 tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc
     4921 caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat
     4981 gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa
     5041 cgacgagcgt gacaccacga tgcctgtagc gatggcaaca acgttgcgca aactattaac
     5101 tggcgaacta cttactctag cttcccggca acaattaata gactggatgg aggcggataa
     5161 agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc
     5221 cggagccggt gagcgtggtt ctcgcggtat catcgcagcg ctggggccag atggtaagcc
     5281 ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag
     5341 acagatcgct gagataggtg cctcactgat taagcattgg taatgaccat gattacgcca
     5401 agctctagct agaggtcgac ggtatacaga catgataaga tacattgatg agtttggaca
     5461 aaccacaact agaatgcagt gaaaaaaatg ctttatttgt gaaatttgtg atgctattgc
     5521 tttatttgta accattataa gctgcaataa acaagttggg gtgggcgaag aactccagca
     5581 tgagatcccc gcgctggagg atcatccagc cggcgtcccg gaaaacgatt ccgaagccca
     5641 acctttcata gaaggcggcg gtggaatcga aatctcgtag cacgtgtcag gcaccgggct
     5701 tgcgggtcat gcaccaggtg cgcggtcctt cgggcacctc gacgtcggcg gtgacggtga
     5761 agccgagccg ctcgtagaag gggaggttgc ggggcgcgga tgtctccagg aaggcgggca
     5821 ccccggcgcg ctcggccgcc tccactccgg ggagcacgac ggcgctgccc agacccttgc
     5881 cctggtggtc gggcgagacg ccgacggtgg ccaggaacca cgcgggctcc ttgggccggt
     5941 gcggcgccag gaggccttcc atctgttgct gcgcggccag ccgggaaccg ctcaactcgg
     6001 ccatgcgcgg gccgatctcg gcgaacaccg cccccgcttc gacgctctcc ggcgtggtcc
     6061 agaccgccac cgcggcgccg tcgtccgcga cccacacctt gccgatgtcg agcccgacgc
     6121 gcgtgaggaa gagttcttgc agctcggtga cccgctcgat gtggcggtcc ggatcgacgg
     6181 tgtggcgcgt ggcggggtag tcggcgaacg cggcggcgag ggtgcgtacg gccctgggga
     6241 cgtcgtcgcg ggtggcgagg cgcaccgtgg gcttgtactc ggtcatggtt tagttcctca
     6301 ccttgtcgta ttatactatg ccgatatact atgccgatga ttaattgtca acacgtgctg
     6361 atcagatccg aaaatggata tacaagctcc cgggagcttt ttgcaaaagc ctaggcctcc
     6421 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcg
//
