LOCUS       Exported                9268 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of cell-surface protein extracellular domain 
            fused to Fc(DAPA)-Avi-6xHis. Protein is secreted from cells..
ACCESSION   .
VERSION     .
KEYWORDS    pD649-HAsp-JAG2-Fc(DAPA)-AviTag-6xHis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9268)
  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 9268)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9268
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     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"
     regulatory      2743..2752
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             6610..6654
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6655..6672
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     primer_bind     complement(6812..6829)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6818..6929
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7047..7067)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7052..7381
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7232..7367
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7294..7313
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7429..7476
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7495..8094
                     /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(7495..7514)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     7991..8011
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8224..8345
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8261..8280)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8315..8334
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(8398..9258)
                     /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     9021..9040
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9259..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 cggccgccat gggatatttc
     2761 gaactgcagc tgagcgccct gcggaacgtt aacggcgaac ttctgtctgg cgcctgctgt
     2821 gatggcgacg gcagaacaac aagagccggc ggatgtggcc acgacgagtg tgatacctac
     2881 gtcagagtgt gcctgaaaga ataccaggcc aaagtgaccc caacaggccc ttgttcttat
     2941 ggccatggcg ccacacctgt gctcggcggc aatagctttt atctgcctcc agctggcgct
     3001 gccggcgata gagctagagc tagggcaaga gctggcggcg atcaagatcc tggcctggtg
     3061 gtcatcccct tccagtttgc ctggcctaga agcttcaccc tgatcgtgga agcctgggac
     3121 tgggacaacg acaccacacc taacgaggaa ctgctgatcg agagagtgtc ccacgccggc
     3181 atgatcaacc ccgaggatag atggaagtcc ctgcacttct ctggacacgt ggcccacctg
     3241 gaactgcaga ttagagtcag atgcgacgag aactactaca gcgccacctg taacaagttc
     3301 tgcagacccc ggaacgactt cttcggccac tacacctgtg accagtacgg caacaaggcc
     3361 tgcatggatg gctggatggg caaagaatgc aaagaggccg tctgcaagca gggctgcaat
     3421 ctgcttcatg gcggctgtac agtgcccggc gagtgcagat gtagctatgg ctggcagggc
     3481 agattctgcg acgagtgcgt tccctatcct ggctgtgtgc acggcagctg tgttgagcct
     3541 tggcagtgca actgcgagac aaactggggc ggactgctgt gcgacaagga cctgaattac
     3601 tgcggcagcc accatccttg caccaacggc ggcacctgta tcaacgccga gccagaccag
     3661 tacagatgca cctgtcctga tggctacagc ggccggaatt gcgagaaagc cgaacacgcc
     3721 tgcaccagca atccttgtgc caatggcgga agctgccacg aggtgccatc tggctttgag
     3781 tgccactgtc ctagcggttg gagcggccct acatgtgccc tggatatcga tgagtgcgcc
     3841 tccaatccat gtgccgctgg tggcacatgt gtggatcagg tggacggctt cgagtgcatc
     3901 tgcccagaac agtgggttgg agccacctgt cagctggacg ccaatgagtg tgaaggcaag
     3961 ccctgcctga acgccttcag ctgcaagaat ctgatcggcg gctactactg cgactgcatc
     4021 cctggctgga agggaatcaa ctgccacatc aacgtgaacg actgcagagg ccagtgtcag
     4081 cacggcggaa catgcaagga tctggtcaac ggctaccagt gcgtgtgccc tagaggattt
     4141 ggcggcagac actgcgagct ggaaagggac gaatgtgcca gctctccttg tcactctggc
     4201 ggcctgtgcg aagatctggc cgatggcttt cactgtcact gccctcaggg cttttctggc
     4261 cctctgtgtg aagtggacgt ggacctgtgt gaaccctctc catgcagaaa cggcgccaga
     4321 tgctacaacc tggaaggcga ctactattgc gcctgtccag acgacttcgg cggaaagaat
     4381 tgcagcgtgc ccagagagcc ttgtcctggc ggagcctgta gagtgatcga tggctgtggc
     4441 tctgatgccg gacctggcat gcctggaaca gctgcttctg gcgtgtgtgg ccctcacgga
     4501 agatgtgttt ctcagcctgg cggcaacttc tcctgcatct gcgatagcgg cttcaccggc
     4561 acctactgtc acgagaacat cgacgactgt ctgggccagc cttgcagaaa tggcggcact
     4621 tgcatcgatg aggtggacgc ctttagatgc ttctgccctt ctggatggga gggcgagctg
     4681 tgcgatacaa accccaacga ctgcctgcct gatccttgcc acagccgggg cagatgttac
     4741 gacctggtca atgacttcta ctgcgcctgc gacgacggat ggaagggcaa gacatgccac
     4801 agcagagaat tccagtgcga cgcctacaca tgtagcaatg gcgggacctg ttacgacagc
     4861 ggcgacacct tcagatgcgc ttgtcctcct ggatggaaag gcagcacttg tgccgtggcc
     4921 aagaacagca gctgtctgcc caatccttgc gtgaacggcg ggacatgtgt tggcagcggc
     4981 gccagcttta gctgtatctg cagagatggc tgggaaggcc ggacatgcac ccacaacacc
     5041 aacgattgca accctctgcc ttgctataac ggcggaatct gcgtggacgg cgtgaactgg
     5101 tttagatgcg agtgcgcccc tggcttcgcc ggaccagact gcagaatcaa cattgacgag
     5161 tgccagagca gcccctgtgc ctacggtgct acttgtgtgg acgagatcaa tggctaccgg
     5221 tgcagctgtc cacctggcag agctggccct agatgccaag aagtgatcgg cttcggcaga
     5281 tcctgctggt ccagaggcac accttttcca cacggctcta gctgggtcga agattgcaac
     5341 agctgcagat gcctggacgg cagacgggac tgcagcaaag tttggtgcgg ctggaagcct
     5401 tgtctgctgg ctggacaacc tgaggctctg tcagcccagt gtcctctggg acagagatgt
     5461 ctggaaaagg cccctggaca gtgcctgagg cctccttgtg aagcttgggg agaatgtggc
     5521 gccgaggaac ctcctagcac accctgtttg cctagatccg gccacctgga caacaactgc
     5581 gccagactga cactgcactt caaccgggat cacgtgccac agggcacaac agtgggagcc
     5641 atctgctccg gaatcagaag cctgcctgcc acaagagctg tggccagaga tagactgctg
     5701 gtgctgctgt gtgatagagc ctcttctggc gcaagcgctg tggaagtggc tgtgtctttt
     5761 agccccgcca gagatctgcc cgacagctct cttattcaag gcgccgctca tgccatcgtg
     5821 gccgccatta cacagagagg caatagcagc ctgctgctgg ccgtgaccga agtgaaggtg
     5881 gaaacagtgg ttaccggcgg cagcagcaca ggcgcgccgg gcggcgacaa gacccacacc
     5941 tgccccccct gcccagctcc agaactgctg ggagggcctt ccgtgttcct gttccccccc
     6001 aagcccaagg acaccctgat gatcagcagg acccccgagg tgacctgcgt ggtggtggcc
     6061 gtgtcccacg aggacccaga ggtgaagttc aactggtacg tggacggcgt ggaggtgcac
     6121 aacgccaaga ccaagcccag agaggagcag tacaacagca cctacagggt ggtgtccgtg
     6181 ctgaccgtgc tgcaccagga ctggctgaac ggcaaagaat acaagtgcaa agtctccaac
     6241 aaggccctgg ctgccccaat cgaaaagaca atcagcaagg ccaagggcca gccacgggag
     6301 ccccaggtgt acaccctgcc ccccagccgg gaggagatga ccaagaacca ggtgtccctg
     6361 acctgtctgg tgaagggctt ctaccccagc gatatcgccg tggagtggga gagcaacggc
     6421 cagcccgaga acaactacaa gaccaccccc ccagtgctgg acagcgacgg cagcttcttc
     6481 ctgtacagca agctgaccgt ggacaagtcc aggtggcagc agggcaacgt gttcagctgc
     6541 agcgtgatgc acgaggccct gcacaaccac tacacccaga agtccctgag cctgagcccc
     6601 ggcggcagcg gtcttaatga tattttcgag gctcagaaga ttgagtggca tgaacaccac
     6661 caccatcacc actaaggccg gcctgtagtc actggcacaa cgcgttaccc ggtaagccaa
     6721 tcgggtatac acggtcgtca tactgcagac agggttcttc tactttgcaa gatagtcttg
     6781 agtagtaaaa taaatagata gagaaaatca gcctcgactg tgccttctag ttgccagcca
     6841 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     6901 ctttcctaat aaaatgagga aattgcatca caacactcaa ccctatctcg gtctattctt
     6961 ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag ctgatttaac
     7021 aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg gaaagtcccc
     7081 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccaggtg
     7141 tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc
     7201 agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc
     7261 ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc
     7321 tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa
     7381 aaagctcccg ggagcttgta tatccatttt cggatctgat cagcacgtgt tgacaattaa
     7441 tcatcggcat agtatatcgg catagtataa tacgacaagg tgaggaacta aaccatgacc
     7501 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     7561 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     7621 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     7681 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     7741 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     7801 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     7861 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     7921 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagaca
     7981 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     8041 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgacacgtg
     8101 ctacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     8161 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     8221 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     8281 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     8341 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatta
     8401 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     8461 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     8521 cgctgcgatg ataccgcgag aaccacgctc accggctccg gatttatcag caataaacca
     8581 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     8641 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     8701 tgttgccatc gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     8761 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     8821 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     8881 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     8941 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     9001 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     9061 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     9121 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     9181 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     9241 gaaatgttga atactcatat tcttcctt
//
