LOCUS       Exported                9376 bp ds-DNA     circular SYN 13-MAY-2021
DEFINITION  Mammalian expression of secreted protein fused to 
            COMP5AP-AviTag-9xHis..
ACCESSION   .
VERSION     .
KEYWORDS    pD649-HAsp-SEMA3D-COMP5AP-AviTag-9xHis
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9376)
  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 9376)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 13, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9376
                     /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"
     CDS             5026..5049
                     /codon_start=1
                     /product="recognition and cleavage site for human 
                     rhinovirus 3C and PreScission proteases"
                     /label=HRV 3C site
                     /translation="LEVLFQGP"
     CDS             6709..6753
                     /codon_start=1
                     /product="peptide tag that allows for enzymatic 
                     biotinylation"
                     /label=AviTag(TM)
                     /translation="GLNDIFEAQKIEWHE"
     CDS             6754..6780
                     /codon_start=1
                     /product="9xHis affinity tag"
                     /label=9xHis
                     /translation="HHHHHHHHH"
     primer_bind     complement(6920..6937)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    6926..7037
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(7155..7175)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        7160..7489
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7340..7475
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     7402..7421
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        7537..7584
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             7603..8202
                     /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(7603..7622)
                     /label=Puro-R
                     /note="Puromycin resistance gene, reverse primer. Also 
                     called puro-variant-R"
     primer_bind     8099..8119
                     /label=Puro-F
                     /note="Puromycin resistance gene, forward primer"
     polyA_signal    8332..8453
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(8369..8388)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     8423..8442
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(8506..9366)
                     /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     9129..9148
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(9367..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 cggccgccac actgaagcag
     2761 aacatcccca gactgaagct gacctacaag gacctgctgc tgagcaacag ctgcatccca
     2821 tttctgggca gcagcgaagg cctggacttt cagacactgc tgctggacga ggaacggggc
     2881 agactgctgc ttggagccaa ggaccacatc tttctgctga gcctggtgga cctgaacaag
     2941 aacttcaaga agatctactg gcctgccgcc aaagaacgcg tggaactgtg taaactggcc
     3001 ggcaaggacg ccaataccga gtgcgccaac ttcatcagag tgctgcagcc ctacaacaag
     3061 acccacatct acgtgtgtgg caccggcgcc ttccatccta tctgcggcta tatcgacctg
     3121 ggcgtgtaca aagaggacat catcttcaag ctggacaccc acaacctgga aagcggcagg
     3181 ctgaagtgcc ccttcgatcc tcagcagcct ttcgccagcg tgatgaccga cgagtacctg
     3241 tacagcggaa ccgccagcga cttcctgggc aaagataccg ccttcaccag atctctgggc
     3301 cccacacacg accaccacta catcagaacc gacatcagcg agcactactg gctgaacggc
     3361 gccaagttta tcggcacctt cttcatcccc gacacctaca atcccgacga cgacaagatc
     3421 tatttcttct tccgggaatc ctctcaagag ggcagcacca gcgacaagac catcctgagc
     3481 agagtgggca gagtgtgcaa gaacgatgtc ggcggacagc ggagcctgat caacaagtgg
     3541 accaccttcc tgaaggcccg gctgatctgt agcatccctg gatctgatgg cgccgatacc
     3601 tacttcgacg agctgcagga tatctacctg ctgcctacca gggacgagag aaaccctgtg
     3661 gtgtacggcg tgttcaccac caccagctcc atctttaagg gcagcgccgt gtgcgtgtac
     3721 agcatggccg atatcagagc cgtgttcaac ggcccctacg ctcacaaaga gtccgccgat
     3781 catagatggg tgcagtacga cggcagaatc ccctatccta gacctggcac atgccccagc
     3841 aagacctacg atcccctgat caagagcacc cgggacttcc ccgatgacgt gatcagcttc
     3901 atcaagcggc actccgtgat gtacaagagc gtgtaccctg tggctggcgg ccctaccttc
     3961 aagagaatca acgtggacta ccggctgacc cagatcgtgg tggatcatgt gatcgccgag
     4021 gacggccagt acgatgtgat gttcctggga acagacatcg gcaccgtgct gaaggtggtg
     4081 tccatcagca aagaaaagtg gaacatggaa gaggtggtgc tcgaggaact gcagatcttt
     4141 aagcacagca gcatcatcct caacatggaa ctgagcctga agcagcagca gctgtacatc
     4201 ggctccagag atggcctggt gcagctgtcc ctgcacagat gtgataccta cggcaaggcc
     4261 tgcgccgatt gctgccttgc cagagatcct tactgtgcct gggatggcaa cgcctgcagc
     4321 agatacgccc caacatctaa gagaagggcc agacggcagg acgtgaagta cggcgatcct
     4381 atcacacagt gctgggacat cgaggacagc atcagccacg agacagccga cgagaaagtg
     4441 atcttcggca tcgagttcaa cagcaccttc ctcgagtgca tccccaagag ccagcaggcc
     4501 accatcaagt ggtacatcca gagaagcggc gacgagcaca gagaggaact gaagccagac
     4561 gagcggatca tcaagaccga gtacggcctg ctgatcagaa gcctgcagaa aaaggacagc
     4621 ggcatgtact actgcaaggc ccaagagcac accttcatcc acaccatcgt gaaactgacc
     4681 ctgaacgtga tcgagaacga gcagatggaa aacacccagc gggccgagca cgaagagggc
     4741 aaagttaagg atctgctcgc cgagagccgg ctgcggtaca aggactatat ccagatcctg
     4801 tctagcccca atttcagcct ggaccagtac tgcgaacaga tgtggcacag agaaaagcgg
     4861 cggcagagaa acaaaggcgg cccaaagtgg aagcacatgc aagagatgaa gaaaaagcgg
     4921 aacagacggc accaccgcga cctggatgaa ctgcctagag ctgtggctac aggcgcgccg
     4981 gaagatcaag ttgatccaag attgattgat ggcaagggcg gatctctgga ggtgctgttc
     5041 cagggaccag gcggaagcgg caatcccggc ggcgatctgg ccccccagat gctgcgcgag
     5101 ctgcaggaga ccaatgccgc cctgcaggac gtgcgcgagc tgctgcgcca ccgcgtgaag
     5161 gagatcacct tcctgaagaa caccgtgatg gagtgcgacg cctgcggtat gcagccctct
     5221 agaggctccg gctccatcat cccagttgag gaggagaacc cggacttctg gaaccgcgag
     5281 gcagccgagg ccctgggtgc cgccaagaag ctgcagcctg cacagacagc cgccaagaac
     5341 ctcatcatct tcctgggcga tgggatgggg gtgtctacgg tgacagctgc caggatccta
     5401 aaagggcaga agaaggacaa actggggcct gagatacccc tggccatgga ccgcttccca
     5461 tatgtggctc tgtccaagac atacaatgta gacaaacatg tgccagacag tggagccaca
     5521 gccacggcct acctgtgcgg ggtcaagggc aacttccaga ccattggctt gagtgcagcc
     5581 gcccgcttta accagtgcaa cacgacacgc ggcaacgagg tcatctccgt gatgaatcgg
     5641 gccaagaaag cagggaagtc agtgggagtg gtaaccacca cacgagtgca gcacgcctcg
     5701 ccagccggca cctacgccca cacggtgaac cgcaactggt actcggacgc cgacgtgcct
     5761 gcctcggccc gccaggaggg gtgccaggac atcgctacgc agctcatctc caacatggac
     5821 attgacgtga tcctaggtgg aggccgaaag tacatgtttc gcatgggaac cccagaccct
     5881 gagtacccag atgactacag ccaaggtggg accaggctgg acgggaagaa tctggtgcag
     5941 gaatggctgg cgaagcgcca gggtgcccgg tatgtgtgga accgcactga gctcatgcag
     6001 gcttccctgg acccgtctgt gacccatctc atgggtctct ttgagcctgg agacatgaaa
     6061 tacgagatcc accgagactc cacactggac ccctccctga tggagatgac agaggctgcc
     6121 ctgcgcctgc tgagcaggaa cccccgcggc ttcttcctct tcgtggaggg tggtcgcatc
     6181 gaccatggtc atcatgaaag cagggcttac cgggcactga ctgagacgat catgttcgac
     6241 gacgccattg agagggcggg ccagctcacc agcgaggagg acacgctgag cctcgtcact
     6301 gccgaccact cccacgtctt ctccttcgga ggctaccccc tgcgagggag ctccatcttc
     6361 gggctggccc ctggcaaggc ccgggacagg aaggcctaca cggtcctcct atacggaaac
     6421 ggtccaggct atgtgctcaa ggacggcgcc cggccggatg ttaccgagag cgagagcggg
     6481 agccccgagt atcggcagca gtcagcagtg cccctggacg aagagaccca cgcaggcgag
     6541 gacgtggcgg tgttcgcgcg cggcccgcag gcgcacctgg ttcacggcgt gcaggagcag
     6601 accttcatag cgcacgtcat ggccttcgcc gcctgcctgg agccctacac cgcctgcgac
     6661 ctggcgcccc ccgccggcac caccgacgcc gcgcacccgg gtggcggagg actgaacgac
     6721 atcttcgagg ctcagaaaat cgaatggcac gaacatcacc atcaccatca ccatcaccat
     6781 taaggccggc ctgtagtcac tggcacaacg cgttacccgg taagccaatc gggtatacac
     6841 ggtcgtcata ctgcagacag ggttcttcta ctttgcaaga tagtcttgag tagtaaaata
     6901 aatagataga gaaaatcagc ctcgactgtg ccttctagtt gccagccatc tgttgtttgc
     6961 ccctcccccg tgccttcctt gaccctggaa ggtgccactc ccactgtcct ttcctaataa
     7021 aatgaggaaa ttgcatcaca acactcaacc ctatctcggt ctattctttt gatttataag
     7081 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     7141 cgaattaatt ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc
     7201 aggcagaagt atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc
     7261 aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt
     7321 cccgccccta actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc
     7381 ccatggctga ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct
     7441 attccagaag tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg
     7501 agcttgtata tccattttcg gatctgatca gcacgtgttg acaattaatc atcggcatag
     7561 tatatcggca tagtataata cgacaaggtg aggaactaaa ccatgaccga gtacaagccc
     7621 acggtgcgcc tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg
     7681 ttcgccgact accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc
     7741 accgagctgc aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc
     7801 gcggacgacg gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg
     7861 gtgttcgccg agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag
     7921 caacagatgg aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc
     7981 accgtcggcg tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc
     8041 ggagtggagg cggccgagcg cgccggggtg cccgccttcc tggagacatc cgcgccccgc
     8101 aacctcccct tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa
     8161 ggaccgcgca cctggtgcat gacccgcaag cccggtgcct gacacgtgct acgagatttc
     8221 gattccaccg ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc
     8281 tggatgatcc tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt
     8341 attgcagctt ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca
     8401 tttttttcac tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc
     8461 tgtataccgt cgacctctag ctagagcttg gcgtaatcat ggtcattacc aatgcttaat
     8521 cagtgaggca cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc
     8581 cgtcgtgtag ataactacga tacgggaggg cttaccatct ggccccagcg ctgcgatgat
     8641 accgcgagaa ccacgctcac cggctccgga tttatcagca ataaaccagc cagccggaag
     8701 ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg
     8761 ccgggaagct agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccatcgc
     8821 tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca
     8881 acgatcaagg cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg
     8941 tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc
     9001 actgcataat tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta
     9061 ctcaaccaag tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc
     9121 aatacgggat aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg
     9181 ttcttcgggg cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc
     9241 cactcgtgca cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc
     9301 aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat
     9361 actcatattc ttcctt
//
