LOCUS       sequence-426817-        7171 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 7171)
  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 7171)
  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-FGFR1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7171
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     124..143
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     149..168
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      254..842
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     743..762
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     996..1013
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     1111..1128
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     1112..1128
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    1584..1600
                     /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        1767..2146
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2147..2350
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     2301..2321
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     2347..2371
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     promoter        2581..2599
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2581..2598
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     2612..2631
                     /label=pMT2-F
                     /note="Synthetic intron, forward primer"
     CDS             4513..4557
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             4558..4575
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(4715..4732)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4721..4832
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4950..4970)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4955..5284
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5135..5270
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5197..5216
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5332..5379
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5398..5997
                     /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(5398..5417)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     5894..5914
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    6127..6248
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6164..6183)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6218..6237
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(6301..7092,7093..7161))
                     /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     6924..6943
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(7162..7171,1..82))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
       61 tgtatttaga aaaataaaca aataggggtc agtgttacaa ccaattaacc aattctgaac
      121 attatcgcga gcccatttat acctgaatat ggctcataac accccttgct catgaccaaa
      181 atcccttaac gtgagttacg cgcgcgtcgt tccactgagc gtcagacccc gtagaaaaga
      241 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
      301 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
      361 aggtaactgg cttcagcaga gcgcagatac caaatactgt tcttctagtg tagccgtagt
      421 tagcccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
      481 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
      541 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
      601 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
      661 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
      721 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
      781 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
      841 aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca
      901 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
      961 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
     1021 aaggcgagag tagggaactg ccaggcatca aactaagcag aaggcccctg acggatggcc
     1081 tttttgcgtt tctacaaact ctttctgtgt tgtaaaacga cggccagtct taagctcggg
     1141 ccccctgggc ggttctgata acgagtaatc gttaatccgc aaataacgta aaaacccgct
     1201 tcggcgggtt tttttatggg gggagtttag ggaaagagca tttgtcagaa tatttaaggg
     1261 cgcctgtcac tttgcttgat atatgagaat tatttaacct tataaatgag aaaaaagcaa
     1321 cgcactttaa ataagatacg ttgctttttc gattgatgaa cacctataat taaactattc
     1381 atctattatt tatgattttt tgtatataca atatttctag tttgttaaag agaattaaga
     1441 aaataaatct cgaaaataat aaagggaaaa tcagtttttg atatcaaaat tatacatgtc
     1501 aacgataata caaaatataa tacaaactat aagatgttat cagtatttat tatcatttag
     1561 aataaatttt gtgtcgccct taattgtgag cggataacaa ttacgagctt catgcacagt
     1621 ggcgttgact caatattggc cattagccat attattcatt ggttatatag cataaatcaa
     1681 tattggctat tggccattgc atacgttgta tctatatcat aatatgtaca tttatattgg
     1741 ctcatgtcca atatgaccgc catgttggca ttgattattg actagttatt aatagtaatc
     1801 aattacgggg tcattagttc atagcccata tatggagttc cgcgttacat aacttacggt
     1861 aaatggcccg cctggctgac cgcccaacga cccccgccca ttgacgtcaa taatgacgta
     1921 tgttcccata gtaacgccaa tagggacttt ccattgacgt caatgggtgg agtatttacg
     1981 gtaaactgcc cacttggcag tacatcaagt gtatcatatg ccaagtacgc cccctattga
     2041 cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag tacatgacct tatgggactt
     2101 tcctacttgg cagtacatct acgtattagt catcgctatt accatggtga tgcggttttg
     2161 gcagtacatc aatgggcgtg gatagcggtt tgactcacgg ggatttccaa gtctccaccc
     2221 cattgacgtc aatgggagtt tgttttggca ccaaaatcaa cgggactttc caaaatgtcg
     2281 taacaactcc gccccattga cgcaaatggg cggtaggcgt gtacggtggg aggtctatat
     2341 aagcagagct cgtttagtga accgtcagat cgcctggaga cgccatccac gctgttttga
     2401 cctccataga agacaccggg accgatccag cctccgcggc cgggaacggt gcattggaac
     2461 gcggattccc cgtgccaaga gtgacgtaag taccgcctat agagtctata ggcccacccc
     2521 cttggcttcg ttagaacgcg gctacaatta atacataacc ttatgtatca tacacatacg
     2581 atttaggtga cactatagaa taacatccac tttgcctttc tctccacagg tgtccactcc
     2641 caggtccaac tgcacctcgg ttctatcgaa aacgcgcctg ctagcgccac catgaagacc
     2701 atcattgccc tgagctacat cttctgcctg gtgttcgctg cggccgccag accttctcct
     2761 acactgcctg aacaggctca gccttgggga gcccctgtgg aagtggaatc ctttctggtg
     2821 caccccggcg atctgctgca gctgagatgc agactgagag atgacgtgca gagcatcaac
     2881 tggctcagag atggggtgca gctggccgag agcaacagaa ccagaatcac cggcgaagag
     2941 gtggaagtgc aggatagcgt gccagccgat tctggcctgt acgcctgtgt gacaagcagc
     3001 cctagcggca gcgataccac ctacttctcc gtgaacgtgt ccgacgctct gcccagcagc
     3061 gaggacgatg acgacgatga tgacagcagc agcgaagaga aagagacaga caacaccaag
     3121 cctaacagaa tgcccgtggc tccctactgg acaagccccg agaagatgga aaagaaactg
     3181 cacgccgtgc ctgccgccaa gaccgtgaag tttaagtgtc ccagctccgg cacacccaat
     3241 ccaacactga gatggctgaa gaacggcaaa gagttcaagc ccgaccaccg gatcggcggc
     3301 tacaaagtcc gatacgccac ctggtccatc atcatggaca gcgtggtgcc cagcgacaag
     3361 ggcaactaca cctgtatcgt ggaaaacgag tacggcagca tcaaccacac ctaccagctg
     3421 gacgtggtgg aaagaagccc tcacagaccc attctgcagg ccggactgcc tgccaacaaa
     3481 acagtggccc tgggcagcaa cgtggaattc atgtgcaagg tgtacagcga ccctcagcct
     3541 cacatccagt ggctgaagca catcgaagtg aacggcagca agatcggccc cgacaacctg
     3601 ccttatgtgc agatcctgaa aacagccggc gtgaacacca ccgacaaaga aatggaagtg
     3661 ctgcacctga ggaatgtgtc cttcgaggac gccggcgagt acacatgtct ggccggcaat
     3721 tctatcggcc tgagccacca ttctgcctgg ctgacagtgc tggaagccct ggaagaaaga
     3781 cccgccgtga tgacaagccc tctgtacctg gaaggcgcgc cgggcggcga caagacccac
     3841 acctgccccc cctgcccagc tccagaactg ctgggagggc cttccgtgtt cctgttcccc
     3901 cccaagccca aggacaccct gatgatcagc aggacccccg aggtgacctg cgtggtggtg
     3961 gccgtgtccc acgaggaccc agaggtgaag ttcaactggt acgtggacgg cgtggaggtg
     4021 cacaacgcca agaccaagcc cagagaggag cagtacaaca gcacctacag ggtggtgtcc
     4081 gtgctgaccg tgctgcacca ggactggctg aacggcaaag aatacaagtg caaagtctcc
     4141 aacaaggccc tggctgcccc aatcgaaaag acaatcagca aggccaaggg ccagccacgg
     4201 gagccccagg tgtacaccct gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc
     4261 ctgacctgtc tggtgaaggg cttctacccc agcgatatcg ccgtggagtg ggagagcaac
     4321 ggccagcccg agaacaacta caagaccacc cccccagtgc tggacagcga cggcagcttc
     4381 ttcctgtaca gcaagctgac cgtggacaag tccaggtggc agcagggcaa cgtgttcagc
     4441 tgcagcgtga tgcacgaggc cctgcacaac cactacaccc agaagtccct gagcctgagc
     4501 cccggcggca gcggtcttaa tgatattttc gaggctcaga agattgagtg gcatgaacac
     4561 caccaccatc accactaagg ccggcctgta gtcactggca caacgcgtta cccggtaagc
     4621 caatcgggta tacacggtcg tcatactgca gacagggttc ttctactttg caagatagtc
     4681 ttgagtagta aaataaatag atagagaaaa tcagcctcga ctgtgccttc tagttgccag
     4741 ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc cactcccact
     4801 gtcctttcct aataaaatga ggaaattgca tcacaacact caaccctatc tcggtctatt
     4861 cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat gagctgattt
     4921 aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt gtggaaagtc
     4981 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     5041 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     5101 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     5161 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     5221 ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc taggcttttg
     5281 caaaaagctc ccgggagctt gtatatccat tttcggatct gatcagcacg tgttgacaat
     5341 taatcatcgg catagtatat cggcatagta taatacgaca aggtgaggaa ctaaaccatg
     5401 accgagtaca agcccacggt gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc
     5461 accctcgccg ccgcgttcgc cgactacccc gccacgcgcc acaccgtcga tccggaccgc
     5521 cacatcgagc gggtcaccga gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc
     5581 ggcaaggtgt gggtcgcgga cgacggcgcc gcggtggcgg tctggaccac gccggagagc
     5641 gtcgaagcgg gggcggtgtt cgccgagatc ggcccgcgca tggccgagtt gagcggttcc
     5701 cggctggccg cgcagcaaca gatggaaggc ctcctggcgc cgcaccggcc caaggagccc
     5761 gcgtggttcc tggccaccgt cggcgtctcg cccgaccacc agggcaaggg tctgggcagc
     5821 gccgtcgtgc tccccggagt ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag
     5881 acatccgcgc cccgcaacct ccccttctac gagcggctcg gcttcaccgt caccgccgac
     5941 gtcgaggtgc ccgaaggacc gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac
     6001 gtgctacgag atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt
     6061 ttccgggacg ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc
     6121 caccccaact tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat
     6181 ttcacaaata aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat
     6241 gtatcttatc atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca
     6301 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6361 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6421 cagcgctgcg atgataccgc gagaaccacg ctcaccggct ccggatttat cagcaataaa
     6481 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6541 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6601 cgttgttgcc atcgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6661 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6721 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6781 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6841 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6901 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     6961 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7021 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7081 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7141 acggaaatgt tgaatactca tattcttcct t
//
