LOCUS       sequence-392665-        8190 bp    DNA     circular SYN 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      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8190)
  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 8190)
  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..8190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(4077..4108)
                     /label=minP
                     /note="minimal TATA-box promoter with low basal activity"
     misc_feature    4287..4378
                     /label=pause site
                     /note="RNA polymerase II transcriptional pause signal from 
                     the human alpha-2 globin gene"
     primer_bind     complement(4319..4338)
                     /label=RVprimer3
                     /note="pGL3 vector, forward primer"
     polyA_signal    4392..4440
                     /note="synthetic polyadenylation signal"
     CDS             join(4545..4613,4614..5405)
                     /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      5605..6193
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6094..6113
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6347..6364
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    6520..6568
                     /note="synthetic polyadenylation signal"
     CDS             complement(6592..7629)
                     /codon_start=1
                     /product="hygromycin B phosphotransferase"
                     /label=HygR
                     /note="confers resistance to hygromycin"
                     /translation="MKKPELTATSVEKFLIEKFDSVSDLMQLSEGEESRAFSFDVGGRG
                     YVLRVNSCADGFYKDRYVYRHFASAALPIPEVLDIGEFSESLTYCISRRAQGVTLQDLP
                     ETELPAVLQPVAEAMDAIAAADLSQTSGFGPFGPQGIGQYTTWRDFICAIADPHVYHWQ
                     TVMDDTVSASVAQALDELMLWAEDCPEVRHLVHADFGSNNVLTDNGRITAVIDWSEAMF
                     GDSQYEVANIFFWRPWLACMEQQTRYFERRHPELAGSPRLRAYMLRIGLDQLYQSLVDG
                     NFDDAAWAQGRCDAIVRSGAGTVGRTQIARRSAAVWTDGCVEVLADSGNRRPSTRPRAK
                     EVGRV"
     RBS             6605..6613
                     /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(7660..8017)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      7674..7809
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(7728..7747)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     enhancer        7826..8017
                     /label=SV40 enhancer
                     /note="enhancer for the SV40 early promoter (Herr, 1993)"
     primer_bind     7974..7994
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
ORIGIN
        1 tgttaaaaac aaaattttgt tgattaataa aagaaataat aaaaagttat gcagacgctg
       61 catggcatta tgcaccatgc ttcctcagaa atacattgag ttttggcgtc cgctgaataa
      121 atggttaata tagtgattat ttggcaacac tgcgcagcat tatgcactgc gcttcctcgg
      181 aaatacgtcg agttttggtg ttgctatatt gcccgcggta agcggggatc taatatacca
      241 gcctgatgca ttatgcacat cagttcctcg gaagtacatc gagttttgct ggtcgggtca
      301 ttggggcgta gcggtcatct tcgtgtgcta gtcagcatca tgctgcaagc aaaaatcata
      361 agtcctataa ttaatagcga cgaggcgccg ccgaaaaggg caaacagcca actccatgaa
      421 gtttttgaga tggcggcttg gaattcttgg tcatttttgt gcggggtgct cacgatatgg
      481 tcagctggtg gtttgcattc tgcattgcat gttgtcttct taccacacag cgatacaata
      541 aagttcgcct gtgggctcgc ggtgctgaag tgtactgtca ccgctccttt ctccaggaca
      601 tgaactgtcg actcttggag ggttgctgtg ctcgaatgcg aatgtacagg gcattgtcct
      661 tcgcggtcgg atacatactg cagggtagcc atcccgccga agtccgctga gtaagtgcac
      721 tcactgacat cacatttgac tgttgagacc agtggtgcat ctgatgtcct gataaaggca
      781 gcgttcggga tgtcgataga gatgggaatg ttcccgtatg agcagtccac cgctcgaagc
      841 ggattgactg caatcttgca cccgaaaggg gcggtttcct gcagtgggcg gcctgagttg
      901 tttttccaca tctcgaatcc agatgcggcc tgcgtgtacg ggacatgcac gttcttggcg
      961 gaaggcttga gtagtctaat gtctgtgctg gcgatgagat ctttgctagt caaggaggta
     1021 gcttgaatgt ctccaaacgc tcctggtttc atcgctccgt attccgggaa gtcatagttg
     1081 tacaccaggc cgcgatggat aacgaccttg tgatcgaatg gtgtaaacga tgctgaaatt
     1141 ggtccagcta tgactttcag gtctttagac gttcctggtg tgactccgtt cacgtacaca
     1201 tctaggaaac tggtagtgtt cccgtacact atacgtagtc ctactttcat cgcggcagta
     1261 tgcaccttaa tcgcctgcgc gtggtcagtc gcgcaatctg ctgacaattc gacgtacgcc
     1321 tcactcatct ggctgttctc actgtcgcaa aaacattgtg ctcctcccca catgaagggg
     1381 tacacccctc caaagacctt gcaggtatag tctgcgtgag cggcgggctg acattccaag
     1441 gagccgcagc atttgacttt aggggagggg accacagtgg tgaatttgca ggtgatgtac
     1501 tcttggttgg tggaaggcaa aacctccgag gacatgacag taatctccaa attgagcggg
     1561 gcgtaccctg ccctttcaac aagtgcctta tacggtatct gtggcacatt tggaacagtg
     1621 gtcgcatgtt cgtaggcgtc taccttcgcc aggtaggcgc cggcaaccac taaaaaaggc
     1681 aggcagcatg agcaacagcg cattagaacg atgacagcgg ccaggggtat acacagctgg
     1741 acccagaaga atggctggct gttcgaccat aggtaactca tggtctcggt gaatgtttca
     1801 gcattagccg acctaacaca gcacaaaagt gccagcgaag ttggaatcac ggcatttggg
     1861 gccagggcat atggcgtcag gcactcacgg cgcgctttac aggcacataa tgctgcaaca
     1921 gttacgccaa tcatcatcgc cacagcagct gatgcgacgg ctaagatggt gtacacagga
     1981 tggcgatggt agtaatgctg tactatttcg tgtggccatc cgtgagggtc tcctggtgca
     2041 gactcttggg catagaccct taccggttcg tgattgcccc atatgtattc caggccatct
     2101 cggtcgacgg tgaagtttct aaccgtcttt ccgatgatcc attcagtagt tggttccgga
     2161 tttgccccta gtctcctggt ggtgagcaat gtcaggtggt ctgtgtctaa ttggaggctg
     2221 atgtgtttaa agccgtgtac tacgttcggc gcgtgggcaa cagggaccat gcaggtactc
     2281 gggatcagct tgaaaggtaa atgcaatttc ccttgggccg tgtggtcggc atgtctgatc
     2341 aagtccggcg aattgaagac ccacttcgtt tggtcgctct tataggcgac gcactgcttg
     2401 atggcggtgc agcccgtgat ttcggtacgg gtcgtaacgg taccggtctt gtaatcgccg
     2461 cacttgcact cgtacgtaat gttctttccg gatggtggct tcgcgtagac tttccctgat
     2521 gattcctcca gataggacgt ataggcgtgc ggtcccggcc tgtgcatagt gatgtagccg
     2581 gcggttgttt ctttcagacg gtcgtacact gtgcaaggaa tcttcttacc gtgaacggga
     2641 ggtaggtcat atttttcccg tcccacgaat tttggtttta tcttgcgggc cattgtgcat
     2701 gacgtcgctg agttgctact cgctatacta accgttacgc tgtcccctgg aggacacttc
     2761 gcgaggagaa agtatccttt gtagctaagc cttctacacg gtcctgaggt gctgatcttg
     2821 atgtcatcca tggtgccttc tttgacagta tgatcctgct cgagcgacat gtagcggtac
     2881 ttatttgagc ttgctgctcc gctttggtcg tatccaaact gggcggaagt ctgtatgcgt
     2941 atggtgttgt cgtccgcttc atcccagacc tgctcgatct taatcgggct aaagcacggt
     3001 tcagtatggt gacagtacga gcatgtgccc aagtacgggc tggtcaaggt aaagtcgtca
     3061 gtgacgcttc ttttacttct gccggacgat ccgcaccgca atatggcgtt gagcagggtg
     3121 tcgtaggcct cgtggttcac gttctcttca aggatgtcaa gagctctgga tggttcgcgg
     3181 gtgtagcatg tgggcgggcg attgcatggg aagctcacgt ttccaagcaa gcacatggcc
     3241 gtgaccagtg gtgctgcaga ccactcttct gtcccttccg gggttgtctt gattgtcttc
     3301 cctttgctat tccaggtgac gaccgaaagg gcagttcttg ttccctcatc agcccctccg
     3361 aggactatcg cgacaacccg gcctgagtta tccataatcg gacgaccact gtctcctctg
     3421 cctcctactc cgcgggggat ggtaaatcta cctccactat actgcaccgc tccgtggtgc
     3481 cagttgtaaa acccttcagg gtgttcgctg gtgtaggtga acgcctcact tctcatgttg
     3541 accggcaact gtgcgaactc catgtcgtat gctgacgact tggtgaattt gagctttgat
     3601 agcacagggt ggtcaatagt tcctttcacg tggagtggtt tcattacctt tccttccatg
     3661 gccagtgcgt gcccgatgac atctccgtcc tcatttttga cgtcgaacag tctgtcggcc
     3721 tccaacttga gtgccatacg ttgtctcttt ccgggtttgg gttttgcagg ttgcttcttc
     3781 ttcttctcct gtgtttttgg tttcttcggc ttcggtggtt gctttggcgc ctgcttcttc
     3841 tggcgcggcg gcgggcgtgg gcgtggggtt tgaggtctag ttgcctgtcc aatgactagg
     3901 gcactgacgg ctgtggtcag ttgctggatt tgggaagcca gcccattgcg ggcaggcatc
     3961 ggggccgcct gcctccttct ccgcggcctc cacatggcag tgggggccgg gaaggggcgg
     4021 cggccgagca tgttaaagaa tcctctattc atggatccca gtaccggatt gccaagctgg
     4081 aagtcgagct tccattatat accctctagt gtctaagctt ggccgccgag gccagatcta
     4141 gatgtcctaa tatggacatc ctagatgtcc taatatggac atcctagatg tcctaatatg
     4201 gacatcctag atgtcctaat atggacatcc tagatgtcct aatatggaca tcctgctagc
     4261 gagctcaggt accggccagt taggccagag aaatgttctg gcacctgcac ttgcactggg
     4321 gacagcctat tttgctagtt tgttttgttt cgttttgttt tgatggagag cgtatgttag
     4381 tactatcgat tcacacaaaa aaccaacaca cagatgtaat gaaaataaag atattttatt
     4441 gcggcctgtc caatacctcc cgtaccttaa tattacttac ttatccttga gagacgtact
     4501 agtaaccctg ataaatgctt caataatatt gaaaaaggac gagtatgagc atccaacatt
     4561 ttcgtgtcgc actcattccc ttctttgcgg cattttgctt gcctgttttt gcacaccccg
     4621 aaacgctggt gaaagtaaaa gatgctgaag atcaactggg tgcaagagtg ggctatatcg
     4681 aactggatct caatagcggc aagatccttg agtctttccg ccccgaagaa cgattcccga
     4741 tgatgagcac tttcaaagta ctgctatgtg gcgcggtgtt gtcccgtata gacgccgggc
     4801 aagagcagct tggtcgccgt atacactact cacaaaacga cttggttgag tactcgccgg
     4861 tcacggaaaa gcatcttacg gatggcatga cggtaagaga attgtgtagt gctgccatta
     4921 ccatgagcga caacaccgcg gccaacttac ttctgacaac gatcggaggc cctaaggagc
     4981 tgactgcatt tcttcataat atgggtgatc atgtgacccg gcttgaccgc tgggaaccag
     5041 agttgaacga agccataccg aacgacgagc gtgataccac gatgccagta gcaatggcca
     5101 caactcttcg gaaactactc actggcgaac ttcttactct agcatcacga cagcagctca
     5161 tagactggat ggaggcggac aaagtagcag gaccacttct tcgctcggcc ctccctgctg
     5221 gctggttcat tgctgacaaa tcgggggccg gtgaacgcgg ctctcgcggc atcattgctg
     5281 cgctggggcc tgatggtaag ccctcacgaa tcgtagtgat ctacacgacg gggagtcagg
     5341 ccactatgga cgaacgaaat aggcagatcg ctgagatcgg tgcctcactg atcaagcact
     5401 ggtaaccact gcagtggttt agcatttgcg gccgctgtca gaccaagttt actcatatat
     5461 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     5521 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     5581 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     5641 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     5701 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg ttcttctagt
     5761 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     5821 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     5881 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     5941 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6001 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6061 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6121 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6181 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6241 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6301 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     6361 cgaggaagcg gaagagcgct gccggcacct gtcctacgag ttgcatgata aagaagacag
     6421 tcataagtgc ggcgacgata gtcatgcccc gcgcccaccg gaaggagctg actgggttga
     6481 aggctctcaa gggcatcggt cgacagcatc tagttcgaac acacaaaaaa ccaacacaca
     6541 gatgtaatga aaataaagat attttattgc ggccatgacc ggttctagag tttaaactcg
     6601 acctacctcc ttagcgcgcg gtcgtgtact gggccggcgg ttgccgctgt cggccagcac
     6661 ctcgacgcag ccgtcggtcc atacggctgc gctccggcga gcgatttgtg tgcgaccgac
     6721 ggtgccggcc ccgctgcgga cgatggcatc gcagcggcct tgagcccagg cagcatcgtc
     6781 gaagttgccg tccacgaggc tctggtagag ctgatccagg ccgatgcgca gcatgtaggc
     6841 tcgcagacga gggctgccgg ccagctcggg atgccggcgc tcgaagtagc gagtctgctg
     6901 ctccatgcaa gccagccagg gccgccagaa gaagatgttg gccacctcgt actgactgtc
     6961 cccgaacata gcttcggacc agtcgattac ggcggtgatg cggccgttgt cggtcaggac
     7021 gttgttgctg ccgaagtcgg catggacgag gtggcgcacc tcgggacagt cttcggccca
     7081 cagcatcagt tcgtccaggg cttgagctac gctggcggac acggtgtcgt ccatcacggt
     7141 ctgccagtgg tagacatggg gatcagcaat ggcgcaaatg aaatcccgcc aggttgtgta
     7201 ctggccgatg ccttggggcc cgaatgggcc gaagccgctg gtttgactga ggtcggcggc
     7261 ggcaatagca tccatagctt cggcgacagg ctgtaacaca gcaggcagct ctgtttcggg
     7321 gaggtcttgg agagtaacgc cttgggcgcg tctactgatg cagtatgtca ggctctcgct
     7381 gaactcgccg atgtccaaca cttcggggat gggtagtgca gcgctggcga agtggcggta
     7441 cacgtagcgg tctttgtaga agccatcagc gcagctattc acccgcagta catagccgcg
     7501 tccgccgaca tcgaagctga aggctcggct ctcttcgccc tccgacaact gcatcaggtc
     7561 gctcacactg tcgaacttct cgatgagaaa tttttcaacg ctggtagcgg tgagttcggg
     7621 cttcttcatg gtggcgctag tgtcagaaga atcgagcttt ttgcaaaagc ctaggcctcc
     7681 aaaaaagcct cctcactact tctggaatag ctcagaggca gaggcggcct cggcctctgc
     7741 ataaataaaa aaaattagtc agccatgggg cggagaatgg gcggaactgg gcggagttag
     7801 gggcgggatg ggcggagtta ggggcgggac tatggttgct gactaattga gatgcatgct
     7861 ttgcatactt ctgcctgctg gggagcctgg ggactttcca cacctggttg ctgactaatt
     7921 gagatgcatg ctttgcatac ttctgcctgc tggggagcct ggggactttc cacaccctaa
     7981 ctgacacaca ttccacagct ggttctttcc gcctcagaag gtagctaacc aagttcctct
     8041 ttcagaggtt atttcaggcc atggtgctgc gcagatcagc ggaagagcgc cccccgggca
     8101 aataggggtt ccgcgcacat ttccccgaaa agtgccacct ggccctttcg tcttcaagaa
     8161 ttaattcccc tcgagctcta gagtttaaac
//
