LOCUS       Exported                8100 bp ds-DNA     circular SYN 11-FEB-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8100)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Friday, Jan 8, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8100
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             complement(455..1315)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /note="This feature has 2 segments:
                        1:  455 .. 1246 / #ccffcc
                        2: 1247 .. 1315 / #ccffcc / signal sequence
                     Cleavage site after base 1246"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(1316..1420)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     LTR             2248..2764
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     misc_feature    2828..3169
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell 
                     virus"
                     /note="color: #ffe4c4"
     CDS             3236..3652
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag 
                     gene lacking the start codon"
                     /note="color: #993366; direction: RIGHT"
     regulatory      3665..3674
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     CDS             3674..3739
                     /codon_start=1
                     /product="three tandem FLAG(R) epitope tags, followed by an
                     enterokinase cleavage site"
                     /label=3xFLAG
                     /note="color: #cc99b2
                     Cleavage site after base 3739"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     CDS             3740..5329
                     /codon_start=1
                     /label=Tox1
                     /note="color: #993366"
                     /translation="LAAAMDVRFYPPPAQPAAAPAAPCLGPSPCLDPYYCNKFDGENMY
                     MSMTEPSQDYVPASQSYPGPSLESEDFNIPPITPPSLPDHSLVHLNEVESGYHSLCHPM
                     NHNGLLPFHPQTMDLPEITVSNMLGQDGALLSNSISVMQEIGNAEGAQYSSHPQMAAMR
                     PRGQPTDIRQQASMMQPGQLTTINQSQLSAQLGLNMGGTNVAHNSPSPPGSKSATPSPS
                     SSVHEDECEDASKINGGEKRPASDMGKKPKTPKKKKKKDPNEPQKPVSAYALFFRDTQA
                     AIKGQNPNATFGEVSKIVASMWDGLGEEQKQVYKKKTEAAKKEYLKQLAAYRASLVSKS
                     YTDPVDVKTSQPPQLVNSKPSVFHGPSQAHSALYLSSHYHQQPGMTPQLTAMHPSLPRN
                     IAPKPNNQMPVTVSIANMAVSPPPPLQISPPLHQHLSMQQHQSLAMQQPLGSQLPMQVQ
                     TALHSPTMQQGFTLQPDYQTIINPTSTAAQVVTQAMEYVRSGCRNPPPQPVDWSTDYCS
                     SGGMQRDKALYLT"
     misc_feature    5357..5909
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
                     /note="color: #a6acb3"
     CDS             5921..6640
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="This feature has 3 segments:
                        1: 5921 .. 5923 / #05fd14
                        2: 5924 .. 5926 / #05fd14 / 1a
                        3: 5927 .. 6640 / #05fd14"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     LTR             6742..7256
                     /label=3' LTR
                     /note="3' long terminal repeat from murine embryonic stem 
                     cell virus"
                     /note="color: #ffe4c4"
     protein_bind    7418..7434
                     /label=lac operator
                     /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)."
                     /note="color: #31849b"
     promoter        complement(7442..7472)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="This reverse directional feature has 3 segments:
                        1: 7442 .. 7448 / #ffffff / -10
                        2: 7449 .. 7466 / #ffffff
                        3: 7467 .. 7472 / #ffffff / -35"
     protein_bind    7487..7508
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(join(7796..8100,1..284))
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
ORIGIN
        1 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
       61 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
      121 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
      181 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
      241 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
      301 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
      361 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
      421 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
      481 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
      541 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
      601 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
      661 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
      721 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
      781 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
      841 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
      901 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
      961 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     1021 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     1081 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     1141 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     1201 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     1261 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     1321 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     1381 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     1441 cacctgacgt ctaagaaacc attattatca tgacattaac ctataaaaat aggcgtatca
     1501 cgaggccctt tcgtctcgcg cgtttcggtg atgacggtga aaacctctga cacatgcagc
     1561 tcccggagac ggtcacagct tgtctgtaag cggatgccgg gagcagacaa gcccgtcagg
     1621 gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa ctatgcggca tcagagcaga
     1681 ttgtactgag agtgcaccat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1741 accgcatcag gcgccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     1801 gggcctcttc gctattacgc cagctggcga aagggggatg tgctgcaagg cgattaagtt
     1861 gggtaacgcc agggttttcc cagtcacgac gttgtaaaac gacggcgcaa ggaatggtgc
     1921 atgcaaggag atggcgccca acagtccccc ggccacgggg cctgccacca tacccacgcc
     1981 gaaacaagcg ctcatgagcc cgaagtggcg agcccgatct tccccatcgg tgatgtcggc
     2041 gatataggcg ccagcaaccg cacctgtggc gccggtgatg ccggccacga tgcgtccggc
     2101 gtagaggcga ttagtccaat ttgttaaaga caggatatca gtggtccagg ctctagtttt
     2161 gactcaacaa tatcaccagc tgaagcctat agagtacgag ccatagataa aataaaagat
     2221 tttatttagt ctccagaaaa aggggggaat gaaagacccc acctgtaggt ttggcaagct
     2281 agcttaagta acgccatttt gcaaggcatg gaaaatacat aactgagaat agagaagttc
     2341 agatcaaggt taggaacaga gagacagcag aatatgggcc aaacaggata tctgtggtaa
     2401 gcagttcctg ccccggctca gggccaagaa cagatggtcc ccagatgcgg tcccgccctc
     2461 agcagtttct agagaaccat cagatgtttc cagggtgccc caaggacctg aaatgaccct
     2521 gtgccttatt tgaactaacc aatcagttcg cttctcgctt ctgttcgcgc gcttctgctc
     2581 cccgagctca ataaaagagc ccacaacccc tcactcggcg cgccagtcct ccgatagact
     2641 gcgtcgcccg ggtacccgta ttcccaataa agcctcttgc tgtttgcatc cgaatcgtgg
     2701 actcgctgat ccttgggagg gtctcctcag attgattgac tgcccacctc gggggtcttt
     2761 catttggagg ttccaccgag atttggagac ccctgcccag ggaccaccga cccccccgcc
     2821 gggaggtaag ctggccagcg gtcgtttcgt gtctgtctct gtctttgtgc gtgtttgtgc
     2881 cggcatctaa tgtttgcgcc tgcgtctgta ctagttagct aactagctct gtatctggcg
     2941 gacccgtggt ggaactgacg agttctgaac acccggccgc aaccctggga gacgtcccag
     3001 ggactttggg ggccgttttt gtggcccgac ctgaggaagg gagtcgatgt ggaatccgac
     3061 cccgtcagga tatgtggttc tggtaggaga cgagaaccta aaacagttcc cgcctccgtc
     3121 tgaatttttg ctttcggttt ggaaccgaag ccgcgcgtct tgtctgctgc agcgctgcag
     3181 catcgttctg tgttgtctct gtctgactgt gtttctgtat ttgtctgaaa attagggcca
     3241 gactgttacc actcccttaa gtttgacctt aggtcactgg aaagatgtcg agcggatcgc
     3301 tcacaaccag tcggtagatg tcaagaagag acgttgggtt accttctgct ctgcagaatg
     3361 gccaaccttt aacgtcggat ggccgcgaga cggcaccttt aaccgagacc tcatcaccca
     3421 ggttaagatc aaggtctttt cacctggccc gcatggacac ccagaccagg tcccctacat
     3481 cgtgacctgg gaagccttgg cttttgaccc ccctccctgg gtcaagccct ttgtacaccc
     3541 taagcctccg cctcctcttc ctccatccgc cccgtctctc ccccttgaac ctcctcgttc
     3601 gaccccgcct cgatcctccc tttatccagc cctcactcct tctctaggcg ccggaattag
     3661 atctgccacc atggactaca aagaccatga cggtgattat aaagatcatg acatcgatta
     3721 caaggatgac gatgacaagc ttgcggccgc gatggacgta agattttatc ctcctccagc
     3781 ccagcccgct gctgcgcccg ccgctccttg tctgggacct tctccctgcc tggaccccta
     3841 ctattgcaac aagtttgacg gggagaacat gtatatgagc atgacagagc cgagccagga
     3901 ctatgtgcca gccagccagt cctaccctgg accaagtctg gaaagtgaag attttaacat
     3961 cccacccatc acgcctccct ccctccctga ccactctctg gtgcacctga atgaggtaga
     4021 atccggttac cactctctgt gtcaccccat gaatcacaat ggcctgctcc cgttccatcc
     4081 acaaaccatg gacctgccag agatcacggt gtccaacatg ctgggccaag atggcgcact
     4141 gctctccaat tccatctctg tgatgcaaga gattgggaac gcagaaggag ctcagtacag
     4201 ctcccatcct cagatggcag ccatgaggcc gagaggccag cctacagaca tcagacagca
     4261 ggcaagcatg atgcagcccg gccagctgac caccatcaac cagtcgcagc tgagcgcaca
     4321 gttgggcttg aacatgggag gaaccaatgt ggcccacaac tccccttctc cacccgggag
     4381 caagtctgca accccttccc catccagctc agtgcatgag gatgagtgtg aggatgcctc
     4441 caagatcaat ggaggagaga agcggcctgc ctctgatatg gggaaaaaac ccaaaactcc
     4501 caagaagaag aagaagaagg atcctaatga gccccagaag cctgtgtctg cctatgcttt
     4561 gttctttcgt gatacccagg ctgccattaa gggccagaat ccaaatgcta cttttggtga
     4621 agtatcgaag attgtggcat caatgtggga tggcctagga gaagaacaga aacaggtcta
     4681 taagaagaaa accgaggctg caaagaagga atacctgaag cagctagcag catacagagc
     4741 cagccttgta tccaagagct acactgatcc tgttgatgtt aagacatctc agccacccca
     4801 gctggtcaac tcaaagccgt cagtattcca tgggcccagc caggcccact cagctctgta
     4861 cctaagttct cactatcacc aacaaccagg aatgactcct cagctcaccg ccatgcatcc
     4921 gagcctcccc agaaacatag cacctaagcc caataaccaa atgccagtga ctgtctccat
     4981 agccaacatg gctgtgtccc caccaccgcc ccttcagatc agcccacctc ttcatcagca
     5041 tctcagcatg cagcagcatc agtcgcttgc catgcagcag ccccttggga gtcagctccc
     5101 catgcaggtc cagactgcct tacactcgcc caccatgcag caaggattta ctcttcaacc
     5161 cgactatcag actattatca accctacatc tacagctgca caagttgtca cccaagcgat
     5221 ggagtacgtg cgttctgggt gcagaaatcc gcccccacag cccgtggact ggagtaccga
     5281 ctactgcagt agtgggggca tgcagaggga caaagcgctg tatctcacct gataagaatt
     5341 ccgccccccc cccctaacgt tactggccga agccgcttgg aataaggccg gtgtgcgttt
     5401 gtctatatgt tattttccac catattgccg tcttttggca atgtgagggc ccggaaacct
     5461 ggccctgtct tcttgacgag cattcctagg ggtctttccc ctctcgccaa aggaatgcaa
     5521 ggtctgttga atgtcgtgaa ggaagcagtt cctctggaag cttcttgaag acaaacaacg
     5581 tctgtagcga ccctttgcag gcagcggaac cccccacctg gcgacaggtg cctctgcggc
     5641 caaaagccac gtgtataaga tacacctgca aaggcggcac aaccccagtg ccacgttgtg
     5701 agttggatag ttgtggaaag agtcaaatgg ctctcctcaa gcgtattcaa caaggggctg
     5761 aaggatgccc agaaggtacc ccattgtatg ggatctgatc tggggcctcg gtgcacatgc
     5821 tttacatgtg tttagtcgag gttaaaaaaa cgtctaggcc ccccgaacca cggggacgtg
     5881 gttttccttt gaaaaacacg atgataatat ggccacaacc atggtgagca agggcgagga
     5941 gctgttcacc ggggtggtgc ccatcctggt cgagctggac ggcgacgtaa acggccacaa
     6001 gttcagcgtg tctggcgagg gcgagggcga tgccacctac ggcaagctga ccctgaagtt
     6061 catctgcacc accggcaagc tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta
     6121 cggcgtgcag tgcttcagcc gctaccccga ccacatgaag cagcacgact tcttcaagtc
     6181 cgccatgccc gaaggctacg tccaggagcg caccatcttc ttcaaggacg acggcaacta
     6241 caagacccgc gccgaggtga agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa
     6301 gggcatcgac ttcaaggagg acggcaacat cctggggcac aagctggagt acaactacaa
     6361 cagccacaac gtctatatca tggccgacaa gcagaagaac ggcatcaagg cgaacttcaa
     6421 gatccgccac aacatcgagg acggcagcgt gcagctcgcc gaccactacc agcagaacac
     6481 ccccatcggc gacggccccg tgctgctgcc cgacaaccac tacctgagca cccagtccgc
     6541 cctgagcaaa gaccccaacg agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc
     6601 cgccgggatc actctcggca tggacgagct gtacaagtaa tgaattaatt aagaattatc
     6661 aagcttatcg ataccgtcga cctgcagcca agcttatcga taaaataaaa gattttattt
     6721 agtctccaga aaaagggggg aatgaaagac cccacctgta ggtttggcaa gctagcttaa
     6781 gtaacgccat tttgcaaggc atggaaaata cataactgag aatagagaag ttcagatcaa
     6841 ggttaggaac agagagacag cagaatatgg gccaaacagg atatctgtgg taagcagttc
     6901 ctgccccggc tcagggccaa gaacagatgg tccccagatg cggtcccgcc ctcagcagtt
     6961 tctagagaac catcagatgt ttccagggtg ccccaaggac ctgaaatgac cctgtgcctt
     7021 atttgaacta accaatcagt tcgcttctcg cttctgttcg cgcgcttctg ctccccgagc
     7081 tcaataaaag agcccacaac ccctcactcg gcgcgccagt cctccgatag actgcgtcgc
     7141 ccgggtaccc gtgtatccaa taaaccctct tgcagttgca tccgacttgt ggtctcgctg
     7201 ttccttggga gggtctcctc tgagtgattg actacccgtc agcgggggtc tttcatgggt
     7261 aacagtttct tgaagttgga gaacaacatt ctgagggtag gagtcgaata ttaagtaatc
     7321 ctgactcaat tagccactgt tttgaatcca catactccaa tactcctgaa atagttcatt
     7381 atggacagcg cagaaagagc tggggagaat tgtgaaattg ttatccgctc acaattccac
     7441 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     7501 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     7561 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     7621 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     7681 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     7741 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     7801 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     7861 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     7921 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     7981 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     8041 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
//
