LOCUS       Exported                5081 bp ds-DNA     circular BCT 08-JAN-2025
DEFINITION  NA 3C flag SA.dna (5079 bp).
ACCESSION   EABR
VERSION     .
KEYWORDS    .
SOURCE      Escherichia coli
  ORGANISM  Escherichia coli
REFERENCE   1  (bases 1 to 5081)
  AUTHORS   Gonzalez-Magaldi M, Gullapalli A, Papoulas O, Liu C, Leung AYH., Guo
            L, Brilot A, Marcotte EM, Ke Z, Leahy DJ.
  TITLE     Structure and organization of full-length Epidermal Growth Factor 
            Receptor in extracellular vesicles by cryo-electron tomography
  JOURNAL   unpublished
REFERENCE   2  (bases 1 to 5081)
  AUTHORS   Gonzalez-Magaldi M, Leahy DJ.
  TITLE     Direct Submission
  JOURNAL   Submitted (09-JAN-2025) Department of Molecular Biosciences, 
            University of Texas at Austin, NHB 4.110, 100 E 24TH ST, Austin, 
            Texas 78712, United States
REFERENCE   3  (bases 1 to 5081)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, Feb 24, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
COMMENT     Bankit Comment: Vecscreen Comment:Submitter says that this sequence 
            is a Synthetic Construct
            Bankit Comment: ALT 
            EMAIL:mgonzalezmagaldi@utexas.edu,dleahy@austin.utexas.edu
            Bankit Comment: TOTAL # OF SEQS:1.
FEATURES             Location/Qualifiers
     source          1..5081
                     /organism="Escherichia coli"
                     /mol_type="genomic DNA"
     source          1..5079
                     /organism="Escherichia coli"
                     /plasmid="EABR NA 3C flag SA"
                     /mol_type="genomic DNA"
                     /label=[cultured bacterial source]
                     /note="[cultured bacterial source]"
                     /db_xref="taxon:562"
     regulatory      96..475
                     /regulatory_class="enhancer"
                     /label=CMV enhancer
                     /note="CMV enhancer"
     regulatory      476..679
                     /regulatory_class="promoter"
                     /label=CMV promoter
                     /note="CMV promoter"
     misc_feature    779..922
                     /label=Fcgr2 (fragment)
                     /note="Fcgr2 (fragment)"
     misc_feature    935..1108
                     /label=CEP55 (fragment)
                     /note="CEP55 (fragment)"
     CDS             1109..1849
                     /codon_start=1
                     /transl_table=11
                     /product="fused protein"
                     /label=fused protein
                     /translation="MNPNQKIITIGSICLVVGLISLILQIGNIISIWISHSIQTGSQNH
                     TGGGGSLEVLFQGPDYKDDDDKDYKDDDDKDYKDDDDKGGGSDPSKDSKAQVSAAEAGI
                     TGTWYNQLGSTFIVTAGADGALTGTYVTARGNAESRYVLTGRYDSAPATDGSGTALGWT
                     VAWKNNYRNAHSATTWSGQYVGGAEARI?TPSGSSRQVQRRQTRGNLHLLVTTRSPKSN
                     RQQQVSMLQRRLG*TMVIRLTQYSS"
     misc_feature    1109..1249
                     /label=NA clone
                     /note="NA clone"
     misc_feature    1127..1212
                     /label=tm
                     /note="tm"
     CDS             1262..1285
                     /codon_start=1
                     /transl_table=11
                     /product="HRV 3C site"
                     /label=HRV 3C site
                     /note="recognition and cleavage site for human rhinovirus
                     3C and PreScission proteases; LEVLFQGP; HRV 3C site"
                     /translation="LEVLFQGP"
     CDS             1286..1333
                     /codon_start=1
                     /transl_table=11
                     /product="2xFLAG"
                     /label=2xFLAG
                     /note="two tandem FLAG epitope tags"
                     /translation="DYKDDDDKDYKDDDDK"
     CDS             1334..1357
                     /codon_start=1
                     /transl_table=11
                     /product="FLAG"
                     /label=FLAG
                     /note="epitope tag followed by an enterokinase cleavage
                     site; DYKDDDDK; FLAG"
                     /translation="DYKDDDDK"
     CDS             1370..1849
                     /codon_start=1
                     /transl_table=11
                     /product="Streptavidin codon optimized 3 mut"
                     /label=Streptavidin codon optimized 3 mut
                     /note="3 mutations for higher afinity to strep tag II like 
                     streptactin"
                     /translation="DPSKDSKAQVSAAEAGITGTWYNQLGSTFIVTAGADGALTGTYVT
                     ARGNAESRYVLTGRYDSAPATDGSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARI
                     ?TPSGSSRQVQRRQTRGNLHLLVTTRSPKSNRQQQVSMLQRRLG*TMVIRLTQYSS"
     regulatory      1881..2105
                     /regulatory_class="polyA_signal_sequence"
                     /note="bGH poly(A) signal; bovine growth hormone
                     polyadenylation signal"
     regulatory      complement(2180..2198)
                     /regulatory_class="promoter"
                     /note="T7 promoter; promoter for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2205..2221)
                     /note="common sequencing primer, one of multiple similar 
                     variants; M13 fwd"
     rep_origin      2363..2818
                     /direction=RIGHT
                     /note="f1 ori; f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand synthesis"
     gene            2844..2948
                     /gene="bla"
                     /label=bla
     regulatory      2844..2948
                     /regulatory_class="promoter"
                     /gene="bla"
                     /gene="bla"
                     /label=AmpR promoter
                     /note="AmpR promoter"
     CDS             2949..3809
                     /codon_start=1
                     /transl_table=11
                     /product="AmpR"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics; beta-lactamase; confers resistance
                     to ampicillin, carbenicillin, and related antibiotics"
                     /translation="M*VFNISVSPLFPFLRHFAFLFLLTQKRW*K*KMLKISWVHEWVT
                     SNWISTAVRSLRVFAPKNVFQ**ALLKFCYVARYYPVLTPGKSNSVAAYTILRMTWLST
                     HQSQKSILRMA*Q*ENYAVLP*P*VITLRPTYF*QRSEDRRS*PLFCTTWGIM*LALIV
                     GNRS*MKPYQTTSVTPRCL*QWQQRCANY*LANYLL*LPGNN**TGWRRIKLQDHFCAR
                     PFRLAGLLLINLEPVSVGLAVSLQHWGQMVSPPVS*LSTRRGVRQLWMNEIDRSLR*VP
                     H*LSIG"
     rep_origin      3980..4568
                     /direction=RIGHT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication; ori"
     protein_bind    4856..4877
                     /label=E. coli catabolite activator protein binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP.; CAP binding site"
     regulatory      4892..4922
                     /regulatory_class="promoter"
                     /note="promoter for the E. coli lac operon; lac promoter"
     protein_bind    4930..4946
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator; 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     4954..4970
                     /note="M13 rev; common sequencing primer, one of multiple 
                     similar variants"
     regulatory      4991..5009
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T3 RNA polymerase; T3 
                     promoter"
ORIGIN
        1 ataaatcaat attggctatt ggccattgca tacgttgtat ccatatcata atatgtacat
       61 ttatattggc tcatgtccaa cattaccgcc atgttgacat tgattattga ctagttatta
      121 atagtaatca attacggggt cattagttca tagcccatat atggagttcc gcgttacata
      181 acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat tgacgtcaat
      241 aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc aatgggtgga
      301 gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc caagtacgcc
      361 ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt acatgacctt
      421 atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta ccatggtgat
      481 gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg gatttccaag
      541 tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac gggactttcc
      601 aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg tacggtggga
      661 ggtctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagac gccatccacg
      721 ctgttttgac ctccatagaa gacaccggga ccgatccagc ctccgcggcc ccaagcttgc
      781 tttgcctggc aatcccgatc acagggaaat gggggaaact ctcccagaag aggttggaga
      841 atatcggcag ccatctggtg gcagtgtacc tgtgtctccg ggtccgccca gtggtcttga
      901 acccactagc tcctcacctt acggcggcgg ctcctttaac agctcaatca acaacatcca
      961 cgaaatggag attcagctga aggatgcact cgaaaagaat caacaatggc tcgtttacga
     1021 ccaacaacgc gaggtgtacg tgaaaggatt gcttgcaaaa attttcgaac tggaaaagaa
     1081 aacagagact gcagcacata gtcttcctat gaacccgaac cagaaaatta ttacgattgg
     1141 atctatctgc cttgtagtcg gtctcatcag cctgatcctc caaataggga atataatctc
     1201 aatatggatc agtcactcaa tacagaccgg cagtcagaat cacaccggag gcggcggatc
     1261 ccttgaagtc ctctttcagg gacccgatta taaggacgat gacgataagg attacaagga
     1321 tgatgatgac aaggattaca aggacgacga tgacaaggga ggcggatccg acccatccaa
     1381 ggactccaaa gcacaagtgt ctgcggctga agccgggata actgggacgt ggtacaacca
     1441 acttgggtct acgtttatag tcactgctgg ggcagacgga gcccttactg gtacctatgt
     1501 gaccgcgaga ggcaacgcag aaagccgcta cgtcctcaca ggtcggtatg atagcgcccc
     1561 cgcgacggat gggagcggga cagcattggg ttggaccgtc gcgtggaaga ataactacag
     1621 aaacgctcat agtgctacca cctggtctgg acagtatgta ggtggagcgg aggcccgaat
     1681 amaaacaccc agtggctcct cacgtcaggt acaacggagg caaacgcgtg gaaatctaca
     1741 cttgttggtc acgacacgtt caccaaagtc aaaccgtcag cagcaagtat cgatgctgca
     1801 aagaaggctg gggtgaacaa tggtaatccg cttgacgcag tacagcagtg agaattcctg
     1861 cagccccgct gatcagcctc gactgtgcct tctagttgcc agccatctgt tgtttgcccc
     1921 tcccccgtgc cttccttgac cctggaaggt gccactccca ctgtcctttc ctaataaaat
     1981 gaggaaattg catcgcattg tctgagtagg tgtcattcta ttctgggggg tggggtgggg
     2041 caggacagca agggggagga ttgggaagac aatagcaggc atgctgggga tgcggtgggc
     2101 tctatggctt ctgaggcgga aagaaccagc tggggctcga tcctctagag cggccgccac
     2161 cgcggtggag ctccaattcg ccctatagtg agtcgtatta caattcactg gccgtcgttt
     2221 tacaacgtcg tgactgggaa aaccctggcg ttacccaact taatcgcctt gcagcacatc
     2281 cccctttcgc cagctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     2341 tgcgcagcct gaatggcgaa tgggacgcgc cctgtagcgg cgcattaagc gcggcgggtg
     2401 tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc gctcctttcg
     2461 ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct ctaaatcggg
     2521 ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa aaacttgatt
     2581 agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc cctttgacgt
     2641 tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca ctcaacccta
     2701 tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat tggttaaaaa
     2761 atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg cttacaattt
     2821 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2881 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2941 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     3001 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     3061 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     3121 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     3181 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     3241 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     3301 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     3361 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     3421 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     3481 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3541 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3601 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3661 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3721 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3781 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3841 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3901 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3961 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     4021 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     4081 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     4141 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     4201 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     4261 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     4321 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     4381 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     4441 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     4501 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4561 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4621 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4681 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4741 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4801 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4861 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4921 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta
     4981 cgccaagctc gaaattaacc ctcactaaag ggaacaaaag ctggtaccgg gccccccctc
     5041 gagatccggc cattagccat attattcatt ggttatatag c
//
