LOCUS       sequence-248641-        6973 bp    DNA     linear   UNA 15-DEC-2025
DEFINITION  Plasmid contains the Sindbis glycoprotein genes and can be converted
            to mRNA for initiating Sindbis virus packaging. See Resource 
            Information section for annotated plasmid sequence..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 6973)
  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 6973)
  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: pSin Helper
FEATURES             Location/Qualifiers
     source          1..6973
                     /mol_type="genomic DNA"
                     /organism="unspecified"
     promoter        complement(5076..5094)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(5077..5094)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     rep_origin      complement(5182..5770)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5262..5281)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5941..6732,6733..6801))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     6564..6583
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6802..6906)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gttaaaaaca aaattttgtt gattaataaa agaaataata aaaagttatg cagacgctgc
       61 atggcattat gcaccatgct tcctcagaaa tacattgagt tttggcgtcc gctgaataaa
      121 tggttaatat agtgattatt tggcaacact gcgcagcatt atgcactgcg cttcctcgga
      181 aatacgtcga gttttggtgt tgctatattg cccgcggtaa gcggggatct aatataccag
      241 cctgatgcat tatgcacatc agttcctcgg aagtacatcg agttttgctg gtcgggtcat
      301 tggggcgtag cggtcatctt cgtgtgctag tcagcatcat gctgcaagca aaaatcataa
      361 gtcctataat taatagcgac gaggcgccgc cgaaaagggc aaacagccaa ctccatgaag
      421 tttttgagat ggcggcttgg aattcttggt catttttgtg cggggtgctc acgatatggt
      481 cagctggtgg tttgcattct gcattgcatg ttgtcttctt accacacagc gatacaataa
      541 agttcgcctg tgggctcgcg gtgctgaagt gtactgtcac cgctcctttc tccaggacat
      601 gaactgtcga ctcttggagg gttgctgtgc tcgaatgcga atgtacaggg cattgtcctt
      661 cgcggtcgga tacatactgc agggtagcca tcccgccgaa gtccgctgag taagtgcact
      721 cactgacatc acatttgact gttgagacca gtggtgcatc tgatgtcctg ataaaggcag
      781 cgttcgggat gtcgatagag atgggaatgt tcccgtatga gcagtccacc gctcgaagcg
      841 gattgactgc aatcttgcac ccgaaagggg cggtttcctg cagtgggcgg cctgagttgt
      901 ttttccacat ctcgaatcca gatgcggcct gcgtgtacgg gacatgcacg ttcttggcgg
      961 aaggcttgag tagtctaatg tctgtgctgg cgatgagatc tttgctagtc aaggaggtag
     1021 cttgaatgtc tccaaacgct cctggtttca tcgctccgta ttccgggaag tcatagttgt
     1081 acaccaggcc gcgatggata acgaccttgt gatcgaatgg tgtaaacgat gctgaaattg
     1141 gtccagctat gactttcagg tctttagacg ttcctggtgt gactccgttc acgtacacat
     1201 ctaggaaact ggtagtgttc ccgtacacta tacgtagtcc tactttcatc gcggcagtat
     1261 gcaccttaat cgcctgcgcg tggtcagtcg cgcaatctgc tgacaattcg acgtacgcct
     1321 cactcatctg gctgttctca ctgtcgcaaa aacattgtgc tcctccccac atgaaggggt
     1381 acacccctcc aaagaccttg caggtatagt ctgcgtgagc ggcgggctga cattccaagg
     1441 agccgcagca tttgacttta ggggagggga ccacagtggt gaatttgcag gtgatgtact
     1501 cttggttggt ggaaggcaaa acctccgagg acatgacagt aatctccaaa ttgagcgggg
     1561 cgtaccctgc cctttcaaca agtgccttat acggtatctg tggcacattt ggaacagtgg
     1621 tcgcatgttc gtaggcgtct accttcgcca ggtaggcgcc ggcaaccact aaaaaaggca
     1681 ggcagcatga gcaacagcgc attagaacga tgacagcggc caggggtata cacagctgga
     1741 cccagaagaa tggctggctg ttcgaccata ggtaactcat ggtctcggtg aatgtttcag
     1801 cattagccga cctaacacag cacaaaagtg ccagcgaagt tggaatcacg gcatttgggg
     1861 ccagggcata tggcgtcagg cactcacggc gcgctttaca ggcacataat gctgcaacag
     1921 ttacgccaat catcatcgcc acagcagctg atgcgacggc taagatggtg tacacaggat
     1981 ggcgatggta gtaatgctgt actatttcgt gtggccatcc gtgagggtct cctggtgcag
     2041 actcttgggc atagaccctt accggttcgt gattgcccca tatgtattcc aggccatctc
     2101 ggtcgacggt gaagtttcta accgtctttc cgatgatcca ttcagtagtt ggttccggat
     2161 ttgcccctag tctcctggtg gtgagcaatg tcaggtggtc tgtgtctaat tggaggctga
     2221 tgtgtttaaa gccgtgtact acgttcggcg cgtgggcaac agggaccatg caggtactcg
     2281 ggatcagctt gaaaggtaaa tgcaatttcc cttgggccgt gtggtcggca tgtctgatca
     2341 agtccggcga attgaagacc cacttcgttt ggtcgctctt ataggcgacg cactgcttga
     2401 tggcggtgca gcccgtgatt tcggtacggg tcgtaacggt accggtcttg taatcgccgc
     2461 acttgcactc gtacgtaatg ttctttccgg atggtggctt cgcgtagact ttccctgatg
     2521 attcctccag ataggacgta taggcgtgcg gtcccggcct gtgcatagtg atgtagccgg
     2581 cggttgtttc tttcagacgg tcgtacactg tgcaaggaat cttcttaccg tgaacgggag
     2641 gtaggtcata tttttcccgt cccacgaatt ttggttttat cttgcgggcc attgtgcatg
     2701 acgtcgctga gttgctactc gctatactaa ccgttacgct gtcccctgga ggacacttcg
     2761 cgaggagaaa gtatcctttg tagctaagcc ttctacacgg tcctgaggtg ctgatcttga
     2821 tgtcatccat ggtgccttct ttgacagtat gatcctgctc gagcgacatg tagcggtact
     2881 tatttgagct tgctgctccg ctttggtcgt atccaaactg ggcggaagtc tgtatgcgta
     2941 tggtgttgtc gtccgcttca tcccagacct gctcgatctt aatcgggcta aagcacggtt
     3001 cagtatggtg acagtacgag catgtgccca agtacgggct ggtcaaggta aagtcgtcag
     3061 tgacgcttct tttacttctg ccggacgatc cgcaccgcaa tatggcgttg agcagggtgt
     3121 cgtaggcctc gtggttcacg ttctcttcga ggatgtcgag agctctggat ggttcgcggg
     3181 tgtagcatgt gggcgggcga ttgcatggga agctcacgtt tccaagcaag cacatggccg
     3241 tgaccagtgg tgcagcagag ggcccagggt tggactcaac gtctcccgcc aacttaagta
     3301 ggtcaaagtt ggcggccgcc tgcttcttct ggcgcggcgg cgggcgtggg cgtggggttt
     3361 gaggtctagt tgcctgtcca atgactaggg cactgacggc tgtggtcagt tgctggattt
     3421 gggaagccag cccattgcgg gcaggcatcg gggccgcctg cctccttctc cgcggcctcc
     3481 acatggcagt gggggctggg aaggggcggc ggccgagcat gttaaagaat cctctattca
     3541 tggtggtggt gttgtggtat tagtcagatg aaatgtacta tgctgactat ttaggaccac
     3601 cgtagagatg ctttatttcc cctctgatgg cttgaaatgc tcttttgctc tgggcaaaag
     3661 ttctcaatgc cagcaggaca ggtgtgatgt tgtctacctc ataccgagtt gccacggcca
     3721 ctgctaaggt gtctgttata cctactctaa accacgcctt tgtttcatct agcagagcgc
     3781 gtcttctgtc ttcgtcttgc tcatcgtcgg ctgggagcgg tttacccaac ttaaacagcc
     3841 ttttcaaggg gtccgccacg cgacacgctg tggaggtaac cgaatcttgc aagatgaatc
     3901 caccgcagaa gtaaggtggt ctctcgccga tgactgcgtc aatgatctta acctccatgt
     3961 tgagccaggt ggcacacctc tcagccattt ctttgtcaga tactactccg tgtataatgt
     4021 tgtcgtcgcc gataaatgct gcacatttgg acgttttaag ccgctcctcc aatactctgc
     4081 tggcgataac gacattcaga actgtgttga caaagagcgt gaggaacatt ccggatttca
     4141 tcatcgcccc gaatttgaaa cgggtacccg tgggcagatg ggtggatgat atttctccaa
     4201 aggcgcactc gatcaagtcg agtagtggtt ggtccacacc caggtcttcc aagatcatca
     4261 ggccggttaa cgccatagcg tcgtcttggc ttttgtcgaa cgaggcgatg ggcttcaact
     4321 ccttcttcct cattatcgac aactttcctg tcctgccttc actcagcttt gtgctgcaga
     4381 gtccgatgtt acgcgcttca aggacttttt cgtcggccca gttggtgttg tatgcagggt
     4441 acgaacctgc catagccgag aacatgaact gggtggtgtc gaagccaatc cagtacaggg
     4501 tccgcacgcc tttcatagcc tggtggtaaa tagttccggg agcgttgatg tacacgtcct
     4561 gcatgacgga gtactcggca cgcgtgttgc aggtaacatc gttgtggaag cagagtgatg
     4621 gcgtttcagc atccggtgta tcaagtacgg tccggaggtc cttgatcttc tcatgcaagt
     4681 tcttgtttgt aatcttacat gctttttccg ccagtttgct ggcatatttc atcatgcggt
     4741 ccgggtcttc tggactacgc atggggcaaa cgcaatggta ctggtgctcg gaaaacattc
     4801 tacgagccgg tgcgctgcct atgtccaaaa tcgtcgctgt ggtaggaacc tccagctcga
     4861 tcagtttact ggccagatgc gaaaatgctc tggcattagc atggtcattt ggagtgacct
     4921 gctgtgctac tacctcaaat tgcgggaagc tcttttgcag ttgcacgaca aacggactct
     4981 gagggtctac gtctacgtta actactggct tctccattgt gatggtagtg caattggtcg
     5041 gctgtttgat tcaatagtgt gtactacgcc gccaatctat agtgtcacct aaatcgtatg
     5101 tgtatgatac ataaggttat gtattaattg tagccgcgtt ctaacgacaa tatgtccata
     5161 cgagctcgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     5221 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     5281 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     5341 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     5401 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     5461 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     5521 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     5581 agagttcttg aagtggtggc ctaactacgg ctacactaga aggacagtat ttggtatctg
     5641 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     5701 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     5761 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     5821 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     5881 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     5941 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6001 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     6061 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     6121 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     6181 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     6241 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     6301 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     6361 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     6421 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     6481 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     6541 ctcttgcccg gcgtcaacac gggataatac cgcgccacat agcagaactt taaaagtgct
     6601 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     6661 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     6721 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     6781 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     6841 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     6901 tccgcgcaca tttccccgaa aagtgccacc tggccctttc gtcttcaaga attaattccc
     6961 ctcgagctct aga
//
