LOCUS       pTSin EGFP               10667 bp ds-DNA     circular SYN 03-JUL-2019
DEFINITION  Plasmid contains the entire replication competent Sindbis genome 
            with the structural genome components replaced by EGFP in an MCS 
            locus..
ACCESSION   .
VERSION     .
KEYWORDS    pTSin EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10667)
  TITLE     Roth Lab Sindbis Plasmids
REFERENCE   2  (bases 1 to 10667)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jul 3, 2019 from SnapGene Server 1.1.58
            //www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..10667
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     regulatory      7682..7691
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             7688..8407
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(7733..7754)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(7994..8013)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     8341..8362
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        8838..8942
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8943..9803
                     /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     complement(9161..9180)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9974..10562
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10463..10482
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     promoter        10650..1
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     10650..10667
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 attggcggcg tagtacacac tattgaatca aacagccgac caattgcact accatcacaa
       61 tggagaagcc agtagttaac gtagacgtag acccgcagag tccgtttgtc gtgcaactgc
      121 aaaagagctt cccgcaattt gaggtagtag cacagcaggt cactccaaat gaccatgcta
      181 atgccagagc attttcgcat ctggccagta aactaatcga gctggaggtt cctaccacag
      241 cgacgatttt ggacataggc agcgcaccgg ctcgtagaat gttttccgag caccagtacc
      301 attgcgtttg ccccatgcgt agtccagaag acccggaccg catgatgaaa tatgccagca
      361 aactggcgga aaaagcatgc aagattacga ataagaactt gcatgagaag atcaaggacc
      421 tccggaccgt acttgataca ccggatgctg aaacgccatc actctgcttc cacaacgatg
      481 ttacctgcaa cacgcgtgcc gagtactccg tcatgcagga cgtgtacatc aacgcacccg
      541 gaactattta ccatcaggct atgaaaggcg tgcggaccct gtactggatt ggcttcgata
      601 ccacccagtt catgttctcg gctatggcag gttcgtaccc tgcgtacaac accaactggg
      661 ccgacgaaaa agtcctcgaa gcgcgtaaca tcggactctg cagcacaaag ctgagtgaag
      721 gcaggacagg aaagttgtcg ataatgagga agaaggagtt gaagcccggg tcacgggttt
      781 atttctccgt tggatcgaca ctttacccag aacacagagc cagcttgcag agctggcatc
      841 ttccatcggt gttccacctg aaaggaaagc agtcgtacac ttgccgctgt gatacagtgg
      901 tgagctgcga aggctacgta gtgaagaaaa tcaccatcag tcccgggatc acgggagaaa
      961 ccgtgggata cgcggttaca aacaatagcg agggcttctt gctatgcaaa gttaccgata
     1021 cagtaaaagg agaacgggta tcgttccccg tgtgcacgta tatcccggcc accatatgcg
     1081 atcagatgac cggcataatg gccacggata tctcacctga cgatgcacaa aaacttctgg
     1141 ttgggctcaa ccagcgaatc gtcattaacg gtaagactaa caggaacacc aataccatgc
     1201 aaaattacct tctgccaatc attgcacaag ggttcagcaa atgggccaag gagcgcaaag
     1261 aagaccttga caatgaaaaa atgctgggta ccagagagcg caagcttaca tatggctgct
     1321 tgtgggcgtt tcgcactaag aaagtgcact cgttctatcg cccacctgga acgcagacca
     1381 tcgtaaaagt cccagcctct tttagcgctt tccccatgtc atccgtatgg actacctctt
     1441 tgcccatgtc gctgaggcag aagataaaat tggcattaca accaaagaag gaggaaaaac
     1501 tgctgcaagt cccggaggaa ttagtcatgg aggccaaggc tgctttcgag gatgctcagg
     1561 aggaatccag agcggagaag ctccgagaag cactcccacc attagtggca gacaaaggta
     1621 tcgaggcagc cgcggaagtt gtctgcgaag tggaggggct ccaggcggac atcggagcag
     1681 cactcgtcga aaccccgcgc ggtcatgtaa ggataatacc tcaagcaaat gaccgtatga
     1741 tcggacagta catcgttgtc tcgccaacct ctgtgctgaa gaacgctaaa ctcgcaccag
     1801 cacacccgct agcagaccag gttaagatca taacgcactc cggaagatca ggaaggtatg
     1861 cagtcgaacc atacgacgct aaagtactga tgccagcagg aagtgccgta ccatggccag
     1921 aattcttagc actgagtgag agcgccacgc tagtgtacaa cgaaagagag tttgtgaacc
     1981 gcaagctgta ccatattgcc atgcacggtc ccgctaagaa tacagaagag gagcagtaca
     2041 aggttacaaa ggcagagctc gcagaaacag agtacgtgtt tgacgtggac aagaagcgat
     2101 gcgtcaagaa ggaagaagcc tcaggacttg tcctctcggg agaactgacc aacccgccct
     2161 atcacgaact agctcttgag ggactgaaga ctcgacccgc ggtcccgtac aaggttgaaa
     2221 caataggagt gataggcaca ccaggatcgg gcaagtcggc tatcatcaag tcaactgtca
     2281 cggcacgtga tcttgttacc agcggaaaga aagaaaactg ccgcgaaatt gaggccgatg
     2341 tgctacggct gaggggcatg cagatcacgt cgaagacagt ggattcggtt atgctcaacg
     2401 gatgccacaa agccgtagaa gtgctgtatg ttgacgaagc gttcgcgtgc cacgcaggag
     2461 cactacttgc cttgattgca atcgtcagac cccgtaagaa ggtagtgcta tgcggagacc
     2521 ctaagcaatg cggattcttc aacatgatgc aactaaaggt atatttcaac cacccggaaa
     2581 aagacatatg taccaagaca ttctacaagt ttatctcccg acgttgcaca cagccagtca
     2641 cggctattgt atcgacactg cattacgatg gaaaaatgaa aaccacaaac ccgtgcaaga
     2701 agaacatcga aatcgacatt acaggggcca cgaagccgaa gccaggggac atcatcctga
     2761 catgcttccg cgggtgggtt aagcaactgc aaatcgacta tcccggacat gaggtaatga
     2821 cagccgcggc ctcacaaggg ctaaccagaa aaggagtata tgccgtccgg caaaaagtca
     2881 atgaaaaccc gctgtacgcg atcacatcag agcatgtgaa cgtgctgctc acccgcactg
     2941 aggacaggct agtatggaaa actttacagg gcgacccatg gattaagcag ctcactaacg
     3001 taccaaaagg aaattttcaa gccaccatcg aggactggga agctgaacac aagggaataa
     3061 ttgctgcgat aaacagtccc gctccccgta ccaatccgtt cagctgcaag actaacgttt
     3121 gctgggcgaa agcactggaa ccgatactgg ccacggccgg tatcgtactt accggttgcc
     3181 agtggagcga gctgttccca cagtttgcag atgacaaacc acactcggcc atctacgccc
     3241 tggacgtaat ctgcattaag tttttcggca tggacttgac aagcggactg ttttccaaac
     3301 agagcatccc gttaacgtac catcctgccg attcagcgag gccagtagct cattgggaca
     3361 acagcccagg aacccgcaag tatgggtacg atcacgccgt tgccgccgaa ctctcccgta
     3421 gatttccggt gttccagcta gctgggaaag gcacacagct tgatttgcag acgggcagaa
     3481 ctagagttat ctccgcacag cataacttgg tcccagtgaa ccgcaatctc ccgcacgcct
     3541 tagtccccga gcacaaggag aaacaacccg gcccggtcaa aaaattcttg agccagttca
     3601 aacaccactc cgtacttgtg gtctcagagg aaaaaattga agctccccac aagagaatcg
     3661 aatggatcgc cccgattggc atagccggcg ctgataagaa ctacaacctg gctttcgggt
     3721 ttccgccgca ggcacggtac gacctggtgt ttatcaatat tggaactaaa tacagaaacc
     3781 atcactttca gcagtgcgaa gaccatgcgg cgaccttgaa aaccctctcg cgttcggccc
     3841 tgaactgcct taaccccgga ggcaccctcg tggtgaagtc ctacggttac gccgaccgca
     3901 atagtgagga cgtagtcacc gctcttgcca gaaaatttgt cagagtgtct gcagcgaggc
     3961 cagagtgcgt ctcaagcaat acagaaatgt acctgatctt ccgacaacta gacaacagcc
     4021 gcacacgaca attcaccccg catcatctga attgtgtgat ttcgtccgtg tacgagggta
     4081 caagagacgg agttggagcc gcaccgtcat accgcactaa aagggagaac attgctgatt
     4141 gtcaagagga agcagttgtc aatgcagcca atccgctggg cagaccaggc gaaggagtct
     4201 gccgtgccat ctataaacgt tggccgaaca gtttcaccga ttcagccaca gagaccggca
     4261 ccgcaaaact gactgtgtgc caaggaaaga aagtgatcca cgcggttggc cctgatttcc
     4321 ggaaacaccc agaggcagaa gccctgaaat tgctgcaaaa cgcctaccat gcagtggcag
     4381 acttagtaaa tgaacataat atcaagtctg tcgccatccc actgctatct acaggcattt
     4441 acgcagccgg aaaagaccgc cttgaagtat cacttaactg cttgacaacc gcgctagata
     4501 gaactgatgc ggacgtaacc atctactgcc tggataagaa gtggaaggaa agaatcgacg
     4561 cggtgctcca acttaaggag tctgtaacag agctgaagga tgaggatatg gagatcgacg
     4621 acgagttagt atggatccat ccggacagtt gcctgaaggg aagaaaggga ttcagtacta
     4681 caaaaggaaa gttgtattcg tactttgaag gcaccaaatt ccatcaagca gcaaaagata
     4741 tggcggagat aaaggtcctg ttcccaaatg accaggaaag caacgagcaa ctgtgtgcct
     4801 acatattggg ggagaccatg gaagcaatcc gcgaaaaatg cccggtcgac cacaacccgt
     4861 cgtctagccc gccaaaaacg ctgccgtgcc tctgcatgta tgccatgacg ccagaaaggg
     4921 tccacagact cagaagcaac aacgtcaaag aagttacagt atgctcctcc accccccttc
     4981 caaagtacaa aatcaagaac gttcagaagg ttcagtgcac aaaagtagtc ctgtttaacc
     5041 cgcatacccc tgcattcgtt cccgcccgta agtacataga agcgccagaa cagcctgcag
     5101 ctccgcctgc acaggccgag gaggcccccg aagttgcagc aacaccaaca ccacctgcag
     5161 ctgataacac ctcgcttgat gtcacggaca tctcactgga catggaagac agtagcgaag
     5221 gctcactctt ttcgagcttt agcggatcgg acaactctat tactagtatg gacagttggt
     5281 cgtcaggacc tagttcacta gagatagtag accgaaggca ggtggtggtg gctgacgtcc
     5341 atgccgtcca agagcctgcc cctgttccac cgccaaggct aaagaagatg gcccgcctgg
     5401 cagcggcaag aatgcaggaa gagccaactc caccggcaag caccagctct gcggacgagt
     5461 cccttcacct ttcttttggt ggggtatcca tgtccttcgg atcccttttc gacggagaga
     5521 tggcccgctt ggcagcggca caacccccgg caagtacatg ccctacggat gtgcctatgt
     5581 ctttcggatc gttttccgac ggagagattg aggagctgag ccgcagagta accgagtctg
     5641 agcccgtcct gtttgggtca tttgaaccgg gcgaagtgaa ctcaattata tcgtcccgat
     5701 cagccgtatc ttttccacca cgcaagcaga gacgtagacg caggagcagg aggaccgaat
     5761 actgactaac cggggtaggt gggtacatat tttcgacgga cacaggccct gggcacttgc
     5821 aaaagaagtc cgttctgcag aaccagctta cagaaccgac cttggagcgc aatgttctgg
     5881 aaagaatcta cgccccggtg ctcgacacgt cgaaagagga acagctcaaa ctcaggtacc
     5941 agatgatgcc caccgaagcc aacaaaagca ggtaccagtc cagaaaagta gaaaatcaga
     6001 aagccataac cactgagcga ctgctttcag ggctacgact gtataactct gccacagatc
     6061 agccagaatg ctataagatc acctacccga aaccatcgta ttccagcagt gtaccggcga
     6121 actactctga cccaaagttt gctgtagctg tttgcaacaa ctatctgcat gagaattacc
     6181 cgacggtagc atcttatcag atcaccgacg agtacgatgc ttacttggat atggtagacg
     6241 ggacagtcgc ttgcctagat actgcaactt tttgccccgc caagcttaga agttacccga
     6301 aaagacacga gtatagagcc ccaaacatcc gcagtgcggt tccatcagcg atgcagaaca
     6361 cgttgcaaaa cgtgctcatt gccgcgacta aaagaaactg caacgtcaca caaatgcgtg
     6421 aattgccaac actggactca gcgacattca acgttgaatg ctttcgaaaa tatgcatgta
     6481 atgacgagta ttgggaggag tttgcccgaa agccaattag gatcactact gagttcgtta
     6541 ccgcatacgt ggccagactg aaaggcccta aggccgccgc actgttcgca aagacgcata
     6601 atttggtccc attgcaagaa gtgcctatgg ataggttcgt catggacatg aaaagagacg
     6661 tgaaagttac acctggcacg aaacacacag aagaaagacc gaaagtacaa gtgatacaag
     6721 ccgcagaacc cctggcgacc gcttacctgt gcgggatcca ccgggagtta gtgcgcaggc
     6781 ttacagccgt cttgctaccc aacattcaca cgctttttga catgtcggcg gaggactttg
     6841 atgcaatcat agcagaacac ttcaagcaag gtgacccggt actggagacg gatatcgcct
     6901 cgttcgacaa aagccaagac gacgctatgg cgttaactgg cctgatgatc ttggaagacc
     6961 tgggtgtgga ccaaccacta ctcgacttga tcgagtgcgc ctttggagaa atatcatcca
     7021 cccatctgcc cacgggtacc cgtttcaaat tcggggcgat gatgaaatcc ggaatgttcc
     7081 tcacgctctt tgtcaacaca gttctgaatg tcgttatcgc cagcagagta ttggaggagc
     7141 ggcttaaaac gtccaaatgt gcagcattta tcggcgacga caacatcata cacggagtag
     7201 tatctgacaa agaaatggct gagaggtgtg ccacctggct caacatggag gttaagatca
     7261 ttgacgcagt catcggcgag agaccgcctt acttctgcgg tggattcatc ttgcaagatt
     7321 cggttacctc cacagcgtgt cgcgtggcgg accccttgaa aaggctgttt aagttgggta
     7381 aaccgctccc agccgacgat gagcaagacg aagacagaag acgcgctctg ctagatgaaa
     7441 caaaggcgtg gtttagagta ggtataacag acaccttagc agtggccgtg gcaactcggt
     7501 atgaggtaga caacatcaca cctgtcctgc tggcattgag aacttttgcc cagagcaaaa
     7561 gagcatttca agccatcaga ggggaaataa agcatctcta cggtggtcct aaatagtcag
     7621 catagtacat ttcatctgac taataccaca acaccaccag cggccgcatc gatcaccggt
     7681 cgccaccatg gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
     7741 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     7801 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     7861 gcccaccctc gtgaccaccc tgacctacgg cgtgcagtgc ttcagccgct accccgacca
     7921 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     7981 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     8041 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     8101 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     8161 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     8221 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     8281 caaccactac ctgagcaccc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     8341 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     8401 caagtaaagc ggcatcgata ctagtacgcc ccaatgaccc gaccagcaaa actcgatgta
     8461 cttccgagga actgatgtgc ataatgcatc aggctggtat attagatccc cgcttaccgc
     8521 gggcaatata gcaacaccaa aactcgacgt atttccgagg aggcgcagtg cataatgctg
     8581 cgcagtgttg ccaaataatc actatattaa ccatttattc agcggacgcc aaaactcaat
     8641 gtatttctga ggaagcatgg tgcataatgc catgcagcgt ctgcataact ttttattatt
     8701 tcttttatta atcaacaaaa ttttgttttt aacatttcaa aaaaaaaaaa aaaaaaaggg
     8761 gatctcgagc tctagagctc gaggggaatt aattcttgaa gacgaaaggg ccaggtggca
     8821 cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata cattcaaata
     8881 tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga
     8941 gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc
     9001 ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg
     9061 cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc
     9121 ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat
     9181 cccgtgttga cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact
     9241 tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat
     9301 tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga
     9361 tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc
     9421 ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga
     9481 tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag
     9541 cttcccggca acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc
     9601 gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt
     9661 ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct
     9721 acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg
     9781 cctcactgat taagcattgg taactgtcag accaagttta ctcatatata ctttagattg
     9841 atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca
     9901 tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga
     9961 tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa
    10021 aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga
    10081 aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt
    10141 taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt
    10201 taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat
    10261 agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct
    10321 tggagcgaac gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca
    10381 cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag
    10441 agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc
    10501 gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga
    10561 aaaacgccag caacgcgagc tcgtatggac atattgtcgt tagaacgcgg ctacaattaa
    10621 tacataacct tatgtatcat acacatacga tttaggtgac actatag
//