LOCUS       Exported                4749 bp ds-DNA     circular BCT 27-JAN-2025
DEFINITION  Escherichia coli strain DH5alpha pBS-CMV-NA-3C-flag-SA clone 
            BS-CMV-NA-3C-flag-SA.
ACCESSION   BSeq#1
VERSION     .
KEYWORDS    .
SOURCE      Escherichia coli
  ORGANISM  Escherichia coli
REFERENCE   1  (bases 1 to 4749)
  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  JOURNAL   unpublished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 4749)
  AUTHORS   Gonzalez-Magaldi M.
  TITLE     Direct Submission
  JOURNAL   Submitted (27-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 4749)
  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: TOTAL # OF SEQS:1
            Bankit Comment: ALT 
            EMAIL:mgonzalezmagaldi@utexas.edu,ldleahy@austin.utexas.edu.
FEATURES             Location/Qualifiers
     source          1..4749
                     /organism="Escherichia coli"
                     /plasmid="pBS-CMV-NA-3C-flag-SA"
                     /strain="DH5alpha"
                     /mol_type="genomic DNA"
                     /isolation_source="bacteria plasmid"
                     /clone="BS-CMV-NA-3C-flag-SA"
                     /label=[cultured bacterial source]
                     /note="[cultured bacterial source]"
                     /db_xref="taxon:562"
     protein_bind    1..4749
                     /label=lac repressor encoded by lacI binding site
                     /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)."
     rep_origin      1..4749
                     /direction=RIGHT
                     /note="f1 ori; f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand synthesis"
     regulatory      96..475
                     /regulatory_class="enhancer"
                     /note="CMV enhancer; human cytomegalovirus immediate early 
                     enhancer"
     regulatory      476..679
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter; CMV promoter"
     CDS             779..919
                     /codon_start=1
                     /transl_table=11
                     /product="Neuraminidase"
                     /label=Neuraminidase
                     /note="Influenza_NA region of fused protein"
                     /translation="MNPNQKIITIGSICLVVGLISLILQIGNIISIWISHSIQTGSQNH
                     TG"
     misc_feature    797..882
                     /label=tm
                     /note="tm"
     CDS             932..955
                     /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; Chain B, cleaved peptide 
                     [synthetic construct]"
                     /translation="LEVLFQGP"
     CDS             956..1027
                     /codon_start=1
                     /transl_table=11
                     /product="3 x FLAG"
                     /label=3 x FLAG
                     /note="tandem FLAG epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     CDS             1040..1519
                     /codon_start=1
                     /transl_table=11
                     /product="Streptavidin"
                     /label=Streptavidin
                     /note="Streptavidin, 3 mutations for higher affinity to
                     strep tag II like streptactin"
                     /translation="DPSKDSKAQVSAAEAGITGTWYNQLGSTFIVTAGADGALTGTYVT
                     ARGNAESRYVLTGRYDSAPATDGSGTALGWTVAWKNNYRNAHSATTWSGQYVGGAEARI
                     NTQWLLTSGTTEANAWKSTLVGHDTFTKVKPSAASIDAAKKAGVNNGNPLDAVQQ"
     regulatory      1551..1775
                     /regulatory_class="polyA_signal_sequence"
                     /note="bovine growth hormone polyadenylation signal; bGH
                     poly(A) signal"
     regulatory      complement(1850..1868)
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase; T7 
                     promoter"
     primer_bind     complement(1875..1891)
                     /note="common sequencing primer, one of multiple similar 
                     variants; M13 fwd"
     gene            2514..2618
                     /gene="bla"
                     /label=bla
     regulatory      2514..2618
                     /regulatory_class="promoter"
                     /gene="bla"
                     /gene="bla"
                     /label=AmpR promoter
                     /note="AmpR promoter"
     gene            2619..3479
                     /gene="bla"
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
     CDS             2619..3479
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="AmpR"
                     /label=bla
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      3650..4238
                     /direction=RIGHT
                     /note="ori; high-copy-number ColE1/pMB1/pBR322/pUC origin
                     of replication"
     protein_bind    4526..4547
                     /label=cAMP binding site
                     /bound_moiety="cAMP"
                     /note="CAP binding activates transcription in the presence
                     of cAMP.; CAP binding site"
     regulatory      4547..4562
                     /regulatory_class="promoter"
                     /note="promoter for the E. coli lac operon; lac promoter"
     primer_bind     4624..4640
                     /note="common sequencing primer, one of multiple similar 
                     variants; M13 rev"
     regulatory      4661..4679
                     /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 ccaagcttat
      781 gaacccgaac cagaaaatta ttacgattgg atctatctgc cttgtagtcg gtctcatcag
      841 cctgatcctc caaataggga atataatctc aatatggatc agtcactcaa tacagaccgg
      901 cagtcagaat cacaccggag gcggcggatc ccttgaagtc ctctttcagg gacccgatta
      961 taaggacgat gacgataagg attacaagga tgatgatgac aaggattaca aggacgacga
     1021 tgacaaggga ggcggatccg acccatccaa ggactccaaa gcacaagtgt ctgcggctga
     1081 agccgggata actgggacgt ggtacaacca acttgggtct acgtttatag tcactgctgg
     1141 ggcagacgga gcccttactg gtacctatgt gaccgcgaga ggcaacgcag aaagccgcta
     1201 cgtcctcaca ggtcggtatg atagcgcccc cgcgacggat gggagcggga cagcattggg
     1261 ttggaccgtc gcgtggaaga ataactacag aaacgctcat agtgctacca cctggtctgg
     1321 acagtatgta ggtggagcgg aggcccgaat aaacacccag tggctcctca cgtcaggtac
     1381 aacggaggca aacgcgtgga aatctacact tgttggtcac gacacgttca ccaaagtcaa
     1441 accgtcagca gcaagtatcg atgctgcaaa gaaggctggg gtgaacaatg gtaatccgct
     1501 tgacgcagta cagcagtgag aattcctgca gccccgctga tcagcctcga ctgtgccttc
     1561 tagttgccag ccatctgttg tttgcccctc ccccgtgcct tccttgaccc tggaaggtgc
     1621 cactcccact gtcctttcct aataaaatga ggaaattgca tcgcattgtc tgagtaggtg
     1681 tcattctatt ctggggggtg gggtggggca ggacagcaag ggggaggatt gggaagacaa
     1741 tagcaggcat gctggggatg cggtgggctc tatggcttct gaggcggaaa gaaccagctg
     1801 gggctcgatc ctctagagcg gccgccaccg cggtggagct ccaattcgcc ctatagtgag
     1861 tcgtattaca attcactggc cgtcgtttta caacgtcgtg actgggaaaa ccctggcgtt
     1921 acccaactta atcgccttgc agcacatccc cctttcgcca gctggcgtaa tagcgaagag
     1981 gcccgcaccg atcgcccttc ccaacagttg cgcagcctga atggcgaatg ggacgcgccc
     2041 tgtagcggcg cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt
     2101 gccagcgccc tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc
     2161 ggctttcccc gtcaagctct aaatcggggg ctccctttag ggttccgatt tagtgcttta
     2221 cggcacctcg accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc
     2281 tgatagacgg tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg
     2341 ttccaaactg gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt
     2401 ttgccgattt cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat
     2461 tttaacaaaa tattaacgct tacaatttag gtggcacttt tcggggaaat gtgcgcggaa
     2521 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     2581 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     2641 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     2701 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     2761 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     2821 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     2881 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     2941 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     3001 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     3061 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     3121 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     3181 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     3241 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     3301 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     3361 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     3421 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     3481 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     3541 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     3601 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     3661 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     3721 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     3781 agataccaaa tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     3841 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     3901 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     3961 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     4021 tgagatacct acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     4081 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     4141 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     4201 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     4261 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     4321 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     4381 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     4441 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     4501 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     4561 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     4621 acacaggaaa cagctatgac catgattacg ccaagctcga aattaaccct cactaaaggg
     4681 aacaaaagct ggtaccgggc cccccctcga gatccggcca ttagccatat tattcattgg
     4741 ttatatagc
//
