LOCUS       Exported                7813 bp ds-DNA     circular BCT 14-JAN-2025
DEFINITION  3C SMO.dna (7813 bp).
ACCESSION   streptagII
VERSION     .
KEYWORDS    .
SOURCE      Escherichia coli
  ORGANISM  Escherichia coli
REFERENCE   1  (bases 1 to 7813)
  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  JOURNAL   unpublished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 7813)
  AUTHORS   Gonzalez-Magaldi M.
  TITLE     Direct Submission
  JOURNAL   Submitted (14-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 7813)
  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..7813
                     /organism="Escherichia coli"
                     /plasmid="streptagII 3C SMO"
                     /mol_type="genomic DNA"
                     /isolation_source="bacteria plasmid"
                     /label=[cultured bacterial source]
                     /note="[cultured bacterial source]"
                     /db_xref="taxon:562"
     regulatory      235..614
                     /regulatory_class="enhancer"
                     /note="human cytomegalovirus immediate early enhancer; CMV 
                     enhancer"
     regulatory      615..818
                     /regulatory_class="promoter"
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter; CMV promoter"
     regulatory      863..881
                     /regulatory_class="promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase; T7 
                     promoter"
     misc_feature    936..941
                     /label=Kozac
                     /note="Kozac"
     misc_feature    1050..1086
                     /label=3c cleaved
                     /note="3c cleaved"
     CDS             1062..1085
                     /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"
                     /translation="LEVLFQGP"
     CDS             1098..3365
                     /codon_start=1
                     /transl_table=11
                     /product="SMO protein"
                     /label=SMO protein
                     /note="7-pass transmembrane receptor Smoothened; SMO"
                     /translation="SSGNATGPGPRSAGGSARRSAAVTGPPPPLSHCGRAAPCEPLRYN
                     VCLGSVLPYGATSTLLAGDSDSQEEAHGKLVLWSGLRNAPRCWAVIQPLLCAVYMPKCE
                     NDRVELPSRTLCQATRGPCAIVERERGWPDFLRCTPDRFPEGCTNEVQNIKFNSSGQCE
                     VPLVRTDNPKSWYEDVEGCGIQCQNPLFTEAEHQDMHSYIAAFGAVTGLCTLFTLATFV
                     ADWRNSNRYPAVILFYVNACFFVGSIGWLAQFMDGARREIVCRADGTMRLGEPTSNETL
                     SCVIIFVIVYYALMAGVVWFVVLTYAWHTSFKALGTTYQPLSGKTSYFHLLTWSLPFVL
                     TVAILAVAQVDGDSVSGICFVGYKNYRYRAGFVLAPIGLVLIVGGYFLIRGVMTLFSIK
                     SNHPGLLSEKAASKINETMLRLGIFGFLAFGFVLITFSCHFYDFFNQAEWERSFRDYVL
                     CQANVTIGLPTKQPIPDCEIKNRPSLLVEKINLFAMFGTGIAMSTWVWTKATLLIWRRT
                     WCRLTGQSDDEPKRIKKSKMIAKAFSKRHELLQNPGQELSFSMHTVSHDGPVAGLAFDL
                     NEPSADVSSAWAQHVTKMVARRGAILPQDISVTPVATPVPPEEQANLWLVEAEISPELQ
                     KRLGRKKKRRKRKKEVCPLAPPPELHPPAPAPSTIPRLPQLPRQKCLVAAGAWGAGDSC
                     RQGAWTLVSNPFCPEPSPPQDPFLPSAPAPVAWAHGRRQGLGPIHSRTNLMDTELMDAD
                     SDF"
     regulatory      3413..3637
                     /regulatory_class="polyA_signal_sequence"
                     /note="bovine growth hormone polyadenylation signal; bGH
                     poly(A) signal"
     rep_origin      3683..4111
                     /direction=RIGHT
                     /note="f1 ori; f1 bacteriophage origin of replication;
                     arrow indicates direction of (+) strand synthesis"
     regulatory      4125..4454
                     /regulatory_class="promoter"
                     /note="SV40 enhancer and early promoter; SV40 promoter"
     rep_origin      4305..4440
                     /note="SV40 ori; SV40 origin of replication"
     gene            4521..5315
                     /gene="aph(3')-II (or nptII)"
                     /label=aph(3')-II (or nptII)
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin)"
     CDS             4521..5315
                     /codon_start=1
                     /transl_table=11
                     /gene="aph(3')-II (or nptII)"
                     /product="NeoR/KanR"
                     /label=aph(3')-II (or nptII)
                     /note="aminoglycoside phosphotransferase from Tn5"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     regulatory      5489..5610
                     /regulatory_class="polyA_signal_sequence"
                     /note="SV40 polyadenylation signal; SV40 poly(A) signal"
     primer_bind     complement(5659..5675)
                     /note="common sequencing primer, one of multiple similar 
                     variants; M13 rev"
     protein_bind    5683..5699
                     /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).; lac 
                     operator"
     regulatory      complement(5707..5737)
                     /regulatory_class="promoter"
                     /note="promoter for the E. coli lac operon; lac promoter"
     protein_bind    5752..5773
                     /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"
     rep_origin      complement(6061..6646)
                     /direction=LEFT
                     /note="ori; high-copy-number ColE1/pMB1/pBR322/pUC origin
                     of replication"
     gene            complement(join(6817..7782,7609..7677))
                     /gene="bla"
                     /label=bla
                     /note="confers resistance to ampicillin, carbenicillin,
                     and related antibiotics"
     CDS             complement(6817..7677)
                     /codon_start=1
                     /transl_table=11
                     /gene="bla"
                     /product="AmpR"
                     /label=bla
                     /note="beta-Lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     regulatory      complement(7678..7782)
                     /regulatory_class="promoter"
                     /gene="bla"
                     /gene="bla"
                     /label=AmpR promoter
                     /note="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 gtttaaactt aagcttggta cccggatcct ctagagccac catgcgaccc tccgggacgg
      961 ccggggcagc gctcctggcg ctgctggctg cgctctgccc ggcgagtcgg gctggcggag
     1021 gcagctggag ccacccgcag ttcgaaaagg gaggaggctc ccttgaagtc ctctttcagg
     1081 gacccggcgg gggctcctcg agcgggaacg cgaccgggcc tgggcctcgg agcgcgggcg
     1141 ggagcgcgag gaggagcgcg gcggtgactg gccctccgcc gccgctgagc cactgcggcc
     1201 gggctgcccc ctgcgagccg ctgcgctaca acgtgtgcct gggctcggtg ctgccctacg
     1261 gggccacctc cacactgctg gccggagact cggactccca ggaggaagcg cacggcaagc
     1321 tcgtgctctg gtcgggcctc cggaatgccc cccgctgctg ggcagtgatc cagcccctgc
     1381 tgtgtgccgt atacatgccc aagtgtgaga atgaccgggt ggagctgccc agccgtaccc
     1441 tctgccaggc cacccgaggc ccctgtgcca tcgtggagag ggagcggggc tggcctgact
     1501 tcctgcgctg cactcctgac cgcttccctg aaggctgcac gaatgaggtg cagaacatca
     1561 agttcaacag ttcaggccag tgcgaagtgc ccttggttcg gacagacaac cccaagagct
     1621 ggtacgagga cgtggagggc tgcggcatcc agtgccagaa cccgctcttc acagaggctg
     1681 agcaccagga catgcacagc tacatcgcgg ccttcggggc cgtcacgggc ctctgcacgc
     1741 tcttcaccct ggccacattc gtggctgact ggcggaactc gaatcgctac cctgctgtta
     1801 ttctcttcta cgtcaatgcg tgcttctttg tgggcagcat tggctggctg gcccagttca
     1861 tggatggtgc ccgccgagag atcgtctgcc gtgcagatgg caccatgagg cttggggagc
     1921 ccacctccaa tgagactctg tcctgcgtca tcatctttgt catcgtgtac tacgccctga
     1981 tggctggtgt ggtttggttt gtggtcctca cctatgcctg gcacacttcc ttcaaagccc
     2041 tgggcaccac ctaccagcct ctctcgggca agacctccta cttccacctg ctcacctggt
     2101 cactcccctt tgtcctcact gtggcaatcc ttgctgtggc gcaggtggat ggggactctg
     2161 tgagtggcat ttgttttgtg ggctacaaga actaccgata ccgtgcgggc ttcgtgctgg
     2221 ccccaatcgg cctggtgctc atcgtgggag gctacttcct catccgagga gtcatgactc
     2281 tgttctccat caagagcaac caccccgggc tgctgagtga gaaggctgcc agcaagatca
     2341 acgagaccat gctgcgcctg ggcatttttg gcttcctggc ctttggcttt gtgctcatta
     2401 ccttcagctg ccacttctac gacttcttca accaggctga gtgggagcgc agcttccggg
     2461 actatgtgct atgtcaggcc aatgtgacca tcgggctgcc caccaagcag cccatccctg
     2521 actgtgagat caagaatcgc ccgagccttc tggtggagaa gatcaacctg tttgccatgt
     2581 ttggaactgg catcgccatg agcacctggg tctggaccaa ggccacgctg ctcatctgga
     2641 ggcgtacctg gtgcaggttg actgggcaga gtgacgatga gccaaagcgg atcaagaaga
     2701 gcaagatgat tgccaaggcc ttctctaagc ggcacgagct cctgcagaac ccaggccagg
     2761 agctgtcctt cagcatgcac actgtgtccc acgacgggcc cgtggcgggc ttggcctttg
     2821 acctcaatga gccctcagct gatgtctcct ctgcctgggc ccagcatgtc accaagatgg
     2881 tggctcggag aggagccata ctgccccagg atatttctgt cacccctgtg gcaactccag
     2941 tgcccccaga ggaacaagcc aacctgtggc tggttgaggc agagatctcc ccagagctgc
     3001 agaagcgcct gggccggaag aagaagagga ggaagaggaa gaaggaggtg tgcccgctgg
     3061 cgccgccccc tgagcttcac ccccctgccc ctgcccccag taccattcct cgactgcctc
     3121 agctgccccg gcagaaatgc ctggtggctg caggtgcctg gggagctggg gactcttgcc
     3181 gacagggagc gtggaccctg gtctccaacc cattctgccc agagcccagt ccccctcagg
     3241 atccatttct gcccagtgca ccggcccccg tggcatgggc tcatggccgc cgacagggcc
     3301 tggggcctat tcactcccgc accaacctga tggacacaga actcatggat gcagactcgg
     3361 acttcttgac tcgagtctag agggcccgtt taaacccgct gatcagcctc gactgtgcct
     3421 tctagttgcc agccatctgt tgtttgcccc tcccccgtgc cttccttgac cctggaaggt
     3481 gccactccca ctgtcctttc ctaataaaat gaggaaattg catcgcattg tctgagtagg
     3541 tgtcattcta ttctgggggg tggggtgggg caggacagca agggggagga ttgggaagac
     3601 aatagcaggc atgctgggga tgcggtgggc tctatggctt ctgaggcgga aagaaccagc
     3661 tggggctcta gggggtatcc ccacgcgccc tgtagcggcg cattaagcgc ggcgggtgtg
     3721 gtggttacgc gcagcgtgac cgctacactt gccagcgccc tagcgcccgc tcctttcgct
     3781 ttcttccctt cctttctcgc cacgttcgcc ggctttcccc gtcaagctct aaatcggggg
     3841 ctccctttag ggttccgatt tagtgcttta cggcacctcg accccaaaaa acttgattag
     3901 ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg tttttcgccc tttgacgttg
     3961 gagtccacgt tctttaatag tggactcttg ttccaaactg gaacaacact caaccctatc
     4021 tcggtctatt cttttgattt ataagggatt ttgccgattt cggcctattg gttaaaaaat
     4081 gagctgattt aacaaaaatt taacgcgaat taattctgtg gaatgtgtgt cagttagggt
     4141 gtggaaagtc cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt
     4201 cagcaaccag gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc
     4261 atctcaatta gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc
     4321 cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg
     4381 ccgaggccgc ctctgcctct gagctattcc agaagtagtg aggaggcttt tttggaggcc
     4441 taggcttttg caaaaagctc ccgggagctt gtatatccat tttcggatct gatcaagaga
     4501 caggatgagg atcgtttcgc atgattgaac aagatggatt gcacgcaggt tctccggccg
     4561 cttgggtgga gaggctattc ggctatgact gggcacaaca gacaatcggc tgctctgatg
     4621 ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct ttttgtcaag accgacctgt
     4681 ccggtgccct gaatgaactg caggacgagg cagcgcggct atcgtggctg gccacgacgg
     4741 gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc gggaagggac tggctgctat
     4801 tgggcgaagt gccggggcag gatctcctgt catctcacct tgctcctgcc gagaaagtat
     4861 ccatcatggc tgatgcaatg cggcggctgc atacgcttga tccggctacc tgcccattcg
     4921 accaccaagc gaaacatcgc atcgagcgag cacgtactcg gatggaagcc ggtcttgtcg
     4981 atcaggatga tctggacgaa gagcatcagg ggctcgcgcc agccgaactg ttcgccaggc
     5041 tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac ccatggcgat gcctgcttgc
     5101 cgaatatcat ggtggaaaat ggccgctttt ctggattcat cgactgtggc cggctgggtg
     5161 tggcggaccg ctatcaggac atagcgttgg ctacccgtga tattgctgaa gagcttggcg
     5221 gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca
     5281 tcgccttcta tcgccttctt gacgagttct tctgagcggg actctggggt tcgaaatgac
     5341 cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg ccttctatga
     5401 aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc agcgcgggga
     5461 tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata atggttacaa
     5521 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     5581 tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga cctctagcta
     5641 gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc cgctcacaat
     5701 tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct aatgagtgag
     5761 ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa acctgtcgtg
     5821 ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc
     5881 ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc
     5941 agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa
     6001 catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt
     6061 tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg
     6121 gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg
     6181 ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag
     6241 cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc
     6301 caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa
     6361 ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg
     6421 taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc
     6481 taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac
     6541 cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggttt
     6601 ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga tctcaagaag atcctttgat
     6661 cttttctacg gggtctgacg ctcagtggaa cgaaaactca cgttaaggga ttttggtcat
     6721 gagattatca aaaaggatct tcacctagat ccttttaaat taaaaatgaa gttttaaatc
     6781 aatctaaagt atatatgagt aaacttggtc tgacagttac caatgcttaa tcagtgaggc
     6841 acctatctca gcgatctgtc tatttcgttc atccatagtt gcctgactcc ccgtcgtgta
     6901 gataactacg atacgggagg gcttaccatc tggccccagt gctgcaatga taccgcgaga
     6961 cccacgctca ccggctccag atttatcagc aataaaccag ccagccggaa gggccgagcg
     7021 cagaagtggt cctgcaactt tatccgcctc catccagtct attaattgtt gccgggaagc
     7081 tagagtaagt agttcgccag ttaatagttt gcgcaacgtt gttgccattg ctacaggcat
     7141 cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc tccggttccc aacgatcaag
     7201 gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat
     7261 cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg gttatggcag cactgcataa
     7321 ttctcttact gtcatgccat ccgtaagatg cttttctgtg actggtgagt actcaaccaa
     7381 gtcattctga gaatagtgta tgcggcgacc gagttgctct tgcccggcgt caatacggga
     7441 taataccgcg ccacatagca gaactttaaa agtgctcatc attggaaaac gttcttcggg
     7501 gcgaaaactc tcaaggatct taccgctgtt gagatccagt tcgatgtaac ccactcgtgc
     7561 acccaactga tcttcagcat cttttacttt caccagcgtt tctgggtgag caaaaacagg
     7621 aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg aaatgttgaa tactcatact
     7681 cttccttttt caatattatt gaagcattta tcagggttat tgtctcatga gcggatacat
     7741 atttgaatgt atttagaaaa ataaacaaat aggggttccg cgcacatttc cccgaaaagt
     7801 gccacctgac gtc
//
