LOCUS       sequence-248622-        8199 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid expresses the Sindbis Structural Genome (SSG) via the serum 
            response element (SRE) promoter. See Resource Information section 
            for annotated plasmid sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 8199)
  AUTHORS   English JG, Olsen RHJ, Lansu K, Patel M, White K, Cockrell AS, Singh
            D, Strachan RT, Wacker D, Roth BL
  TITLE     VEGAS as a Platform for Facile Directed Evolution in Mammalian 
            Cells.
  JOURNAL   Cell. 2019 Jul 2. pii: S0092-8674(19)30622-1. doi: 
            10.1016/j.cell.2019.05.051.
  PUBMED    31280962
REFERENCE   2  (bases 1 to 8199)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Cell. 2019 
            Jul 2. pii: S0092-8674(19)30622-1. doi: 10.1016/j.cell.2019.05.051."
COMMENT     Sequence Label: pSRE SSG
FEATURES             Location/Qualifiers
     source          1..8199
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(4086..4117)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    4296..4387
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4328..4347)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4401..4449
                     /note="synthetic polyadenylation signal"
     CDS             join(4554..4622,4623..5414)
                     /codon_start=1
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5614..6202
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6103..6122
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6356..6373
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6529..6577
                     /note="synthetic polyadenylation signal"
     CDS             complement(6601..7638)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6614..6622
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(7669..8026)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7683..7818
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7737..7756)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        7835..8026
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7983..8003
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 tttttgaaat gttaaaaaca aaattttgtt gattaataaa agaaataata aaaagttatg
       61 cagacgctgc atggcattat gcaccatgct tcctcagaaa tacattgagt tttggcgtcc
      121 gctgaataaa tggttaatat agtgattatt tggcaacact gcgcagcatt atgcactgcg
      181 cttcctcgga aatacgtcga gttttggtgt tgctatattg cccgcggtaa gcggggatct
      241 aatataccag cctgatgcat tatgcacatc agttcctcgg aagtacatcg agttttgctg
      301 gtcgggtcat tggggcgtag cggtcatctt cgtgtgctag tcagcatcat gctgcaagca
      361 aaaatcataa gtcctataat taatagcgac gaggcgccgc cgaaaagggc aaacagccaa
      421 ctccatgaag tttttgagat ggcggcttgg aattcttggt catttttgtg cggggtgctc
      481 acgatatggt cagctggtgg tttgcattct gcattgcatg ttgtcttctt accacacagc
      541 gatacaataa agttcgcctg tgggctcgcg gtgctgaagt gtactgtcac cgctcctttc
      601 tccaggacat gaactgtcga ctcttggagg gttgctgtgc tcgaatgcga atgtacaggg
      661 cattgtcctt cgcggtcgga tacatactgc agggtagcca tcccgccgaa gtccgctgag
      721 taagtgcact cactgacatc acatttgact gttgagacca gtggtgcatc tgatgtcctg
      781 ataaaggcag cgttcgggat gtcgatagag atgggaatgt tcccgtatga gcagtccacc
      841 gctcgaagcg gattgactgc aatcttgcac ccgaaagggg cggtttcctg cagtgggcgg
      901 cctgagttgt ttttccacat ctcgaatcca gatgcggcct gcgtgtacgg gacatgcacg
      961 ttcttggcgg aaggcttgag tagtctaatg tctgtgctgg cgatgagatc tttgctagtc
     1021 aaggaggtag cttgaatgtc tccaaacgct cctggtttca tcgctccgta ttccgggaag
     1081 tcatagttgt acaccaggcc gcgatggata acgaccttgt gatcgaatgg tgtaaacgat
     1141 gctgaaattg gtccagctat gactttcagg tctttagacg ttcctggtgt gactccgttc
     1201 acgtacacat ctaggaaact ggtagtgttc ccgtacacta tacgtagtcc tactttcatc
     1261 gcggcagtat gcaccttaat cgcctgcgcg tggtcagtcg cgcaatctgc tgacaattcg
     1321 acgtacgcct cactcatctg gctgttctca ctgtcgcaaa aacattgtgc tcctccccac
     1381 atgaaggggt acacccctcc aaagaccttg caggtatagt ctgcgtgagc ggcgggctga
     1441 cattccaagg agccgcagca tttgacttta ggggagggga ccacagtggt gaatttgcag
     1501 gtgatgtact cttggttggt ggaaggcaaa acctccgagg acatgacagt aatctccaaa
     1561 ttgagcgggg cgtaccctgc cctttcaaca agtgccttat acggtatctg tggcacattt
     1621 ggaacagtgg tcgcatgttc gtaggcgtct accttcgcca ggtaggcgcc ggcaaccact
     1681 aaaaaaggca ggcagcatga gcaacagcgc attagaacga tgacagcggc caggggtata
     1741 cacagctgga cccagaagaa tggctggctg ttcgaccata ggtaactcat ggtctcggtg
     1801 aatgtttcag cattagccga cctaacacag cacaaaagtg ccagcgaagt tggaatcacg
     1861 gcatttgggg ccagggcata tggcgtcagg cactcacggc gcgctttaca ggcacataat
     1921 gctgcaacag ttacgccaat catcatcgcc acagcagctg atgcgacggc taagatggtg
     1981 tacacaggat ggcgatggta gtaatgctgt actatttcgt gtggccatcc gtgagggtct
     2041 cctggtgcag actcttgggc atagaccctt accggttcgt gattgcccca tatgtattcc
     2101 aggccatctc ggtcgacggt gaagtttcta accgtctttc cgatgatcca ttcagtagtt
     2161 ggttccggat ttgcccctag tctcctggtg gtgagcaatg tcaggtggtc tgtgtctaat
     2221 tggaggctga tgtgtttaaa gccgtgtact acgttcggcg cgtgggcaac agggaccatg
     2281 caggtactcg ggatcagctt gaaaggtaaa tgcaatttcc cttgggccgt gtggtcggca
     2341 tgtctgatca agtccggcga attgaagacc cacttcgttt ggtcgctctt ataggcgacg
     2401 cactgcttga tggcggtgca gcccgtgatt tcggtacggg tcgtaacggt accggtcttg
     2461 taatcgccgc acttgcactc gtacgtaatg ttctttccgg atggtggctt cgcgtagact
     2521 ttccctgatg attcctccag ataggacgta taggcgtgcg gtcccggcct gtgcatagtg
     2581 atgtagccgg cggttgtttc tttcagacgg tcgtacactg tgcaaggaat cttcttaccg
     2641 tgaacgggag gtaggtcata tttttcccgt cccacgaatt ttggttttat cttgcgggcc
     2701 attgtgcatg acgtcgctga gttgctactc gctatactaa ccgttacgct gtcccctgga
     2761 ggacacttcg cgaggagaaa gtatcctttg tagctaagcc ttctacacgg tcctgaggtg
     2821 ctgatcttga tgtcatccat ggtgccttct ttgacagtat gatcctgctc gagcgacatg
     2881 tagcggtact tatttgagct tgctgctccg ctttggtcgt atccaaactg ggcggaagtc
     2941 tgtatgcgta tggtgttgtc gtccgcttca tcccagacct gctcgatctt aatcgggcta
     3001 aagcacggtt cagtatggtg acagtacgag catgtgccca agtacgggct ggtcaaggta
     3061 aagtcgtcag tgacgcttct tttacttctg ccggacgatc cgcaccgcaa tatggcgttg
     3121 agcagggtgt cgtaggcctc gtggttcacg ttctcttcaa ggatgtcaag agctctggat
     3181 ggttcgcggg tgtagcatgt gggcgggcga ttgcatggga agctcacgtt tccaagcaag
     3241 cacatggccg tgaccagtgg tgctgcagac cactcttctg tcccttccgg ggttgtcttg
     3301 attgtcttcc ctttgctatt ccaggtgacg accgaaaggg cagttcttgt tccctcatca
     3361 gcccctccga ggactatcgc gacaacccgg cctgagttat ccataatcgg acgaccactg
     3421 tctcctctgc ctcctactcc gcgggggatg gtaaatctac ctccactata ctgcaccgct
     3481 ccgtggtgcc agttgtaaaa cccttcaggg tgttcgctgg tgtaggtgaa cgcctcactt
     3541 ctcatgttga ccggcaactg tgcgaactcc atgtcgtatg ctgacgactt ggtgaatttg
     3601 agctttgata gcacagggtg gtcaatagtt cctttcacgt ggagtggttt cattaccttt
     3661 ccttccatgg ccagtgcgtg cccgatgaca tctccgtcct catttttgac gtcgaacagt
     3721 ctgtcggcct ccaacttgag tgccatacgt tgtctctttc cgggtttggg ttttgcaggt
     3781 tgcttcttct tcttctcctg tgtttttggt ttcttcggct tcggtggttg ctttggcgcc
     3841 tgcttcttct ggcgcggcgg cgggcgtggg cgtggggttt gaggtctagt tgcctgtcca
     3901 atgactaggg cactgacggc tgtggtcagt tgctggattt gggaagccag cccattgcgg
     3961 gcaggcatcg gggccgcctg cctccttctc cgcggcctcc acatggcagt gggggccggg
     4021 aaggggcggc ggccgagcat gttaaagaat cctctattca tggatcccag taccggattg
     4081 ccaagctgga agtcgagctt ccattatata ccctctagtg tctaagcttg gccgccgagg
     4141 ccagatctag atgtcctaat atggacatcc tagatgtcct aatatggaca tcctagatgt
     4201 cctaatatgg acatcctaga tgtcctaata tggacatcct agatgtccta atatggacat
     4261 cctgctagcg agctcaggta ccggccagtt aggccagaga aatgttctgg cacctgcact
     4321 tgcactgggg acagcctatt ttgctagttt gttttgtttc gttttgtttt gatggagagc
     4381 gtatgttagt actatcgatt cacacaaaaa accaacacac agatgtaatg aaaataaaga
     4441 tattttattg cggcctgtcc aatacctccc gtaccttaat attacttact tatccttgag
     4501 agacgtacta gtaaccctga taaatgcttc aataatattg aaaaaggacg agtatgagca
     4561 tccaacattt tcgtgtcgca ctcattccct tctttgcggc attttgcttg cctgtttttg
     4621 cacaccccga aacgctggtg aaagtaaaag atgctgaaga tcaactgggt gcaagagtgg
     4681 gctatatcga actggatctc aatagcggca agatccttga gtctttccgc cccgaagaac
     4741 gattcccgat gatgagcact ttcaaagtac tgctatgtgg cgcggtgttg tcccgtatag
     4801 acgccgggca agagcagctt ggtcgccgta tacactactc acaaaacgac ttggttgagt
     4861 actcgccggt cacggaaaag catcttacgg atggcatgac ggtaagagaa ttgtgtagtg
     4921 ctgccattac catgagcgac aacaccgcgg ccaacttact tctgacaacg atcggaggcc
     4981 ctaaggagct gactgcattt cttcataata tgggtgatca tgtgacccgg cttgaccgct
     5041 gggaaccaga gttgaacgaa gccataccga acgacgagcg tgataccacg atgccagtag
     5101 caatggccac aactcttcgg aaactactca ctggcgaact tcttactcta gcatcacgac
     5161 agcagctcat agactggatg gaggcggaca aagtagcagg accacttctt cgctcggccc
     5221 tccctgctgg ctggttcatt gctgacaaat cgggggccgg tgaacgcggc tctcgcggca
     5281 tcattgctgc gctggggcct gatggtaagc cctcacgaat cgtagtgatc tacacgacgg
     5341 ggagtcaggc cactatggac gaacgaaata ggcagatcgc tgagatcggt gcctcactga
     5401 tcaagcactg gtaaccactg cagtggttta gcatttgcgg ccgctgtcag accaagttta
     5461 ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa
     5521 gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc
     5581 gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat
     5641 ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga
     5701 gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt
     5761 tcttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata
     5821 cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac
     5881 cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg
     5941 ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg
     6001 tgagctatga gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag
     6061 cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct
     6121 ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc
     6181 aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt
     6241 ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg
     6301 tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga
     6361 gtcagtgagc gaggaagcgg aagagcgctg ccggcacctg tcctacgagt tgcatgataa
     6421 agaagacagt cataagtgcg gcgacgatag tcatgccccg cgcccaccgg aaggagctga
     6481 ctgggttgaa ggctctcaag ggcatcggtc gacagcatct agttcgaaca cacaaaaaac
     6541 caacacacag atgtaatgaa aataaagata ttttattgcg gccatgaccg gttctagagt
     6601 ttaaactcga cctacctcct tagcgcgcgg tcgtgtactg ggccggcggt tgccgctgtc
     6661 ggccagcacc tcgacgcagc cgtcggtcca tacggctgcg ctccggcgag cgatttgtgt
     6721 gcgaccgacg gtgccggccc cgctgcggac gatggcatcg cagcggcctt gagcccaggc
     6781 agcatcgtcg aagttgccgt ccacgaggct ctggtagagc tgatccaggc cgatgcgcag
     6841 catgtaggct cgcagacgag ggctgccggc cagctcggga tgccggcgct cgaagtagcg
     6901 agtctgctgc tccatgcaag ccagccaggg ccgccagaag aagatgttgg ccacctcgta
     6961 ctgactgtcc ccgaacatag cttcggacca gtcgattacg gcggtgatgc ggccgttgtc
     7021 ggtcaggacg ttgttgctgc cgaagtcggc atggacgagg tggcgcacct cgggacagtc
     7081 ttcggcccac agcatcagtt cgtccagggc ttgagctacg ctggcggaca cggtgtcgtc
     7141 catcacggtc tgccagtggt agacatgggg atcagcaatg gcgcaaatga aatcccgcca
     7201 ggttgtgtac tggccgatgc cttggggccc gaatgggccg aagccgctgg tttgactgag
     7261 gtcggcggcg gcaatagcat ccatagcttc ggcgacaggc tgtaacacag caggcagctc
     7321 tgtttcgggg aggtcttgga gagtaacgcc ttgggcgcgt ctactgatgc agtatgtcag
     7381 gctctcgctg aactcgccga tgtccaacac ttcggggatg ggtagtgcag cgctggcgaa
     7441 gtggcggtac acgtagcggt ctttgtagaa gccatcagcg cagctattca cccgcagtac
     7501 atagccgcgt ccgccgacat cgaagctgaa ggctcggctc tcttcgccct ccgacaactg
     7561 catcaggtcg ctcacactgt cgaacttctc gatgagaaat ttttcaacgc tggtagcggt
     7621 gagttcgggc ttcttcatgg tggcgctagt gtcagaagaa tcgagctttt tgcaaaagcc
     7681 taggcctcca aaaaagcctc ctcactactt ctggaatagc tcagaggcag aggcggcctc
     7741 ggcctctgca taaataaaaa aaattagtca gccatggggc ggagaatggg cggaactggg
     7801 cggagttagg ggcgggatgg gcggagttag gggcgggact atggttgctg actaattgag
     7861 atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac acctggttgc
     7921 tgactaattg agatgcatgc tttgcatact tctgcctgct ggggagcctg gggactttcc
     7981 acaccctaac tgacacacat tccacagctg gttctttccg cctcagaagg tagctaacca
     8041 agttcctctt tcagaggtta tttcaggcca tggtgctgcg cagatcagcg gaagagcgcc
     8101 ccccgggcaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg gccctttcgt
     8161 cttcaagaat taattcccct cgagctctag agtttaaac
//
