LOCUS       sequence-401039-        7237 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 7237)
  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 7237)
  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-ROR1-Fc(DAPA)-AviTag-6xHis
FEATURES             Location/Qualifiers
     source          1..7237
                     /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             2813..2857
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             2858..2875
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(3015..3032)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3021..3132
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(3250..3270)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3255..3584
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3435..3570
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3497..3516
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        3632..3679
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             3698..4297
                     /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(3698..3717)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     4194..4214
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    4427..4548
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4464..4483)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4518..4537
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(4601..5392,5393..5461))
                     /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     5224..5243
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5462..5553)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5595..5614
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
     primer_bind     5620..5639
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     rep_origin      5725..6313
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6214..6233
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6467..6484
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     6582..6599
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     6583..6599
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    7055..7071
                     /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 cccaagaaac agagctgtct gtgtctgccg agctggtgcc
     1021 taccagcagc tggaatatca gcagcgagct gaacaaggac agctacctga cactggacga
     1081 gcccatgaac aacatcacca ccagcctggg acagaccgcc gagctgcact gtaaagtgtc
     1141 cggcaatcct cctcctacca tccggtggtt caagaacgat gcccctgtgg tgcaagagcc
     1201 cagacggctg agcttcagaa gcaccatcta cggcagcaga ctgcggatca gaaacctgga
     1261 caccaccgac accggctact tccagtgtgt ggccaccaac ggcaaagaag tggtgtctag
     1321 caccggcgtg ctgttcgtga agtttggccc tcctccaacc gcctctcctg gctactctga
     1381 cgagtacgaa gaggacggct tctgccagcc ttaccgggga attgcctgcg ccagattcat
     1441 cggcaaccgg accgtgtaca tggaaagcct gcacatgcag ggcgagatcg agaaccagat
     1501 caccgccgcc tttaccatga tcggcacaag cagccacctg agcgacaagt gcagccagtt
     1561 cgccattcct agcctgtgcc actacgcctt tccatactgc gacgagacaa gcagcgtgcc
     1621 caagcctaga gatctgtgca gagatgagtg cgagatcctg gaaaacgtgc tgtgccagac
     1681 cgagtacatc ttcgcccgca gcaaccccat gatcctgatg agactgaagc tgcccaactg
     1741 cgaggacctg cctcagcctg aatctcctga agccgccaat tgcatccgga tcggcatccc
     1801 tatggccgat cctatcaaca agaaccacaa gtgctacaac agcacaggcg tggactacag
     1861 aggcaccgtg tccgtgacaa agtccggcag acagtgccag ccatggaaca gccagtatcc
     1921 tcacacacac accttcacag ccctgcggtt ccctgagctg aatggcggcc acagctactg
     1981 cagaaacccc ggcaatcaga aagaggcccc ttggtgcttc accctggacg agaacttcaa
     2041 gagcgacctg tgcgacatcc ccgcctgcga tagcaaggac tccaaagaaa agaacaagat
     2101 ggaaatcctg tatggcgcgc cgggcggcga caagacccac acctgccccc cctgcccagc
     2161 tccagaactg ctgggagggc cttccgtgtt cctgttcccc cccaagccca aggacaccct
     2221 gatgatcagc aggacccccg aggtgacctg cgtggtggtg gccgtgtccc acgaggaccc
     2281 agaggtgaag ttcaactggt acgtggacgg cgtggaggtg cacaacgcca agaccaagcc
     2341 cagagaggag cagtacaaca gcacctacag ggtggtgtcc gtgctgaccg tgctgcacca
     2401 ggactggctg aacggcaaag aatacaagtg caaagtctcc aacaaggccc tggctgcccc
     2461 aatcgaaaag acaatcagca aggccaaggg ccagccacgg gagccccagg tgtacaccct
     2521 gccccccagc cgggaggaga tgaccaagaa ccaggtgtcc ctgacctgtc tggtgaaggg
     2581 cttctacccc agcgatatcg ccgtggagtg ggagagcaac ggccagcccg agaacaacta
     2641 caagaccacc cccccagtgc tggacagcga cggcagcttc ttcctgtaca gcaagctgac
     2701 cgtggacaag tccaggtggc agcagggcaa cgtgttcagc tgcagcgtga tgcacgaggc
     2761 cctgcacaac cactacaccc agaagtccct gagcctgagc cccggcggca gcggtcttaa
     2821 tgatattttc gaggctcaga agattgagtg gcatgaacac caccaccatc accactaagg
     2881 ccggcctgta gtcactggca caacgcgtta cccggtaagc caatcgggta tacacggtcg
     2941 tcatactgca gacagggttc ttctactttg caagatagtc ttgagtagta aaataaatag
     3001 atagagaaaa tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc
     3061 ccccgtgcct tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga
     3121 ggaaattgca tcacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     3181 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     3241 taattctgtg gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca
     3301 gaagtatgca aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct
     3361 ccccagcagg cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc
     3421 ccctaactcc gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg
     3481 gctgactaat tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc
     3541 agaagtagtg aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt
     3601 gtatatccat tttcggatct gatcagcacg tgttgacaat taatcatcgg catagtatat
     3661 cggcatagta taatacgaca aggtgaggaa ctaaaccatg accgagtaca agcccacggt
     3721 gcgcctcgcc acccgcgacg acgtccccag ggccgtacgc accctcgccg ccgcgttcgc
     3781 cgactacccc gccacgcgcc acaccgtcga tccggaccgc cacatcgagc gggtcaccga
     3841 gctgcaagaa ctcttcctca cgcgcgtcgg gctcgacatc ggcaaggtgt gggtcgcgga
     3901 cgacggcgcc gcggtggcgg tctggaccac gccggagagc gtcgaagcgg gggcggtgtt
     3961 cgccgagatc ggcccgcgca tggccgagtt gagcggttcc cggctggccg cgcagcaaca
     4021 gatggaaggc ctcctggcgc cgcaccggcc caaggagccc gcgtggttcc tggccaccgt
     4081 cggcgtctcg cccgaccacc agggcaaggg tctgggcagc gccgtcgtgc tccccggagt
     4141 ggaggcggcc gagcgcgccg gggtgcccgc cttcctggag acatccgcgc cccgcaacct
     4201 ccccttctac gagcggctcg gcttcaccgt caccgccgac gtcgaggtgc ccgaaggacc
     4261 gcgcacctgg tgcatgaccc gcaagcccgg tgcctgacac gtgctacgag atttcgattc
     4321 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4381 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact tgtttattgc
     4441 agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata aagcattttt
     4501 ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc atgtctgtat
     4561 accgtcgacc tctagctaga gcttggcgta atcatggtca ttaccaatgc ttaatcagtg
     4621 aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg
     4681 tgtagataac tacgatacgg gagggcttac catctggccc cagcgctgcg atgataccgc
     4741 gagaaccacg ctcaccggct ccggatttat cagcaataaa ccagccagcc ggaagggccg
     4801 agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg
     4861 aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc atcgctacag
     4921 gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat
     4981 caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc
     5041 cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc
     5101 ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa
     5161 ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac
     5221 gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt
     5281 cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc
     5341 gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa
     5401 caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca
     5461 tattcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat
     5521 acatatttga atgtatttag aaaaataaac aaataggggt cagtgttaca accaattaac
     5581 caattctgaa cattatcgcg agcccattta tacctgaata tggctcataa caccccttgc
     5641 tcatgaccaa aatcccttaa cgtgagttac gcgcgcgtcg ttccactgag cgtcagaccc
     5701 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5761 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5821 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5881 gtagccgtag ttagcccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5941 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6001 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6061 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6121 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6181 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6241 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6301 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6361 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6421 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6481 cgaggaagcg gaaggcgaga gtagggaact gccaggcatc aaactaagca gaaggcccct
     6541 gacggatggc ctttttgcgt ttctacaaac tctttctgtg ttgtaaaacg acggccagtc
     6601 ttaagctcgg gccccctggg cggttctgat aacgagtaat cgttaatccg caaataacgt
     6661 aaaaacccgc ttcggcgggt ttttttatgg ggggagttta gggaaagagc atttgtcaga
     6721 atatttaagg gcgcctgtca ctttgcttga tatatgagaa ttatttaacc ttataaatga
     6781 gaaaaaagca acgcacttta aataagatac gttgcttttt cgattgatga acacctataa
     6841 ttaaactatt catctattat ttatgatttt ttgtatatac aatatttcta gtttgttaaa
     6901 gagaattaag aaaataaatc tcgaaaataa taaagggaaa atcagttttt gatatcaaaa
     6961 ttatacatgt caacgataat acaaaatata atacaaacta taagatgtta tcagtattta
     7021 ttatcattta gaataaattt tgtgtcgccc ttaattgtga gcggataaca attacgagct
     7081 tcatgcacag tggcgttgac tcaatattgg ccattagcca tattattcat tggttatata
     7141 gcataaatca atattggcta ttggccattg catacgttgt atctatatca taatatgtac
     7201 atttatattg gctcatgtcc aatatgaccg ccatgtt
//
