LOCUS       Exported                8933 bp ds-DNA     circular SYN 13-APR-2018
DEFINITION  synthetic circular DNA
ACCESSION   urn.local...1367881019035.10
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8933)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 12, 2018 from SnapGene 4.0.8
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..8933
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     812..831
                     /label=BII(R)
     repeat_region   887..917
                     /label=5'TR(1)
     misc_feature    887..917
                     /label=31bp spacer
     enhancer        1112..1343
                     /label=core insul enhancer
                     /label=core insul
     enhancer        1406..2132
                     /label=CMV promoter (with TetOs)
     promoter        1406..2033
                     /label=CMV
                     /note="CMV"
     misc_feature    1891
                     /label=A
                     /note="Geneious type: Editing History Replacement"
     TATA_signal     1978..1984
     misc_feature    1994..2033
                     /label=TetR binding sites
                     /note="TetR binding sites"
     misc_feature    1994..2012
                     /label=TetO
                     /note="TetO"
     misc_feature    2015..2033
                     /label=TetO
                     /note="TetO"
     misc_feature    2156..2163
                     /label=Kozak
                     /note="Kozak"
     primer_bind     2156..2163
                     /label=primer3
                     /note="Geneious name: Primer"
     CDS             2167..2199
                     /codon_start=1
                     /label=Lyn
                     /translation="GCIKSKGKDSA"
     CDS             2212..2232
                     /codon_start=1
                     /label=TVMVs
                     /translation="ETVRFQY"
     CDS             2233..2274
                     /codon_start=1
                     /label=KDegron
                     /translation="HKSGAWKLPVSLVK"
     CDS             2287..2928
                     /codon_start=1
                     /label=SpGFP1-10
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATIGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     KYKTRAVVKFEGDTLVNRIELKGTDFKEDGNILGHKLEYNFNSHNVYITADKQKNGIKA
                     NFTVRHNVEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQTVLSKDPNEK"
     CDS             2944..3510
                     /codon_start=1
                     /label=CmIFP
                     /translation="TTNIAPALDGAFHRITSSSSLMGLCDETATIIREITGYDRVMVVR
                     FDEEGNAEILSERRRADLEAFLGNRYPASTIPQIARRLYEHNRVRLLVDVNYTPVPLQP
                     RISPLNGRDLDMSLSCLRSMSPVHQKYMQDMGVGATLVCSLMVSGRLWGLIACHHYEPR
                     FVPFHIRAAGEALAETCANRIATLES"
     CDS             3511..3531
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQS"
     CDS             3538..3858
                     /codon_start=1
                     /label=NmIFP
                     /translation="CEREQIHLAGSIQPHGILLAVKEPDNVVIQASINAAEFLNTNSVV
                     GRPLRDLGGDLPLQILPHLNGPLHLAPVTLRCTVGSPPRRVDCTIHRPSNGGLIVELEP
                     ATK"
     CDS             3871..3921
                     /codon_start=1
                     /label=SpGFP11
                     /translation="SRDHMVLHEYVNAAGIT"
     polyA_signal    3938..4152
                     /label=BGH polyadenylation sequence
     misc_feature    4226..4551
                     /label=SV40 Promoter
     misc_feature    4230
                     /label=G missing
     CDS             4587..5381
                     /codon_start=1
                     /label=neo
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     misc_feature    5382..5387
                     /label=SalI
                     /note="Geneious type: restriction site"
     misc_feature    5388..5492
                     /label=WPRE
     misc_feature    join(5498..5501,5506,5508..5633)
                     /label=SV40 pA
     enhancer        complement(5882..6112)
                     /label=core insul(1) enhancer
                     /label=core insul(1)
     repeat_region   6387..6421
                     /label=3'TR
     primer_bind     complement(6940..6959)
                     /label=pLac(F)
     CDS             complement(8076..8933)
                     /label=Amp
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccacta gtaggcgttt tgcgctgctt
     1381 cgcgatgtac gggccagata tacgcgttga cattgattat tgactagtta ttaatagtaa
     1441 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
     1501 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
     1561 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
     1621 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
     1681 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
     1741 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
     1801 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
     1861 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
     1921 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
     1981 ataagcagag ctctccctat cagtgataga gatctcccta tcagtgatag agatcgtcga
     2041 cgagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg ttttgacctc
     2101 catagaagac accgggaccg atccagcctc cggactctag cgtttaaact taagctcgcc
     2161 accatggggt gcataaagtc aaaggggaaa gactctgcag gaagcttgag tgaaacagtt
     2221 cgcttccaat accacaagag cggtgcttgg aaacttcccg tctctttggt aaagggagga
     2281 acctcaatgt ccaaaggaga agaactgttt accggtgttg tgccaatttt ggttgaactc
     2341 gatggtgatg tcaacggaca taagttctca gtgagaggcg aaggagaagg tgacgccacc
     2401 attggaaaat tgactcttaa attcatctgt actactggta aacttcctgt accatggccg
     2461 actctcgtaa caacgcttac gtacggagtt cagtgctttt cgagataccc agaccatatg
     2521 aaaagacatg acttttttaa gtcggctatg cctgaaggtt acgtgcaaga aagaacaatt
     2581 tcgttcaaag atgatggaaa atataaaact agagcagttg ttaaatttga aggagatact
     2641 ttggttaacc gcattgaact gaaaggaaca gattttaaag aagatggtaa tattcttgga
     2701 cacaaactcg aatacaattt taatagtcat aacgtataca tcactgctga taagcaaaag
     2761 aacggaatta aagcgaattt cacagtacgc cataatgtag aagatggcag tgttcaactt
     2821 gccgaccatt accaacaaaa cacccctatt ggagacggtc cggtacttct tcctgataat
     2881 cactacctct caacacaaac agtcctgagc aaagatccaa atgaaaaagg aggcagcgga
     2941 tccaccacta acattgcgcc ggctctggac ggtgcgtttc atcgtatcac ttcttcatcc
     3001 tccctgatgg gcctgtgtga cgaaaccgcg actattatcc gtgagattac tggctacgac
     3061 cgtgtgatgg tagtacgttt cgatgaagag ggtaatgccg aaattctgtc cgaacgtcgt
     3121 cgtgcggacc tggaagcgtt cctgggtaac cgctacccgg cgtctactat tccgcagatc
     3181 gctcgtcgcc tgtacgaaca taaccgtgtt cgcctgctgg tagatgtgaa ctatactccg
     3241 gttccgctac agccgcgcat cagcccgctg aacggtcgtg atctggatat gtccctgtct
     3301 tgcctgcgct ctatgtcccc ggtccaccag aaatacatgc aggacatggg cgttggcgcg
     3361 accctggttt gctctctgat ggtgtctggt cgtttgtggg gtctgatcgc ttgccaccac
     3421 tacgaaccgc gcttcgttcc gttccacatt cgcgctgctg gcgaagcgct ggcggaaact
     3481 tgtgcgaacc gcatcgcgac gctggagagc gaaaatctct acttccagag tgaattctgt
     3541 gaacgcgagc agatccacct ggcgggctcc attcagccgc acggtatcct gctggctgtg
     3601 aaagagccag acaacgtggt gatccaggct tctatcaacg ctgcggagtt cctgaacacc
     3661 aactctgttg ttggccgtcc gctgcgtgac ctgggcggcg atctgccttt gcagatcctg
     3721 ccgcacctga acggcccgct gcacctggct ccggtgaccc tgcgttgtac cgtgggttct
     3781 ccgccgcgcc gtgtggactg taccattcac cgtccgtcta acggcggcct gatcgtagaa
     3841 ctggaaccag caaccaagct cgagggaggc agccgggacc acatggtgct gcacgagtac
     3901 gtgaacgccg ccggcatcac atagcgaatt taaatcggcc tcgactgtgc cttctagttg
     3961 ccagccatct gttgtttgcc cctcccccgt gccttccttg accctggaag gtgccactcc
     4021 cactgtcctt tcctaataaa atgaggaaat tgcatcgcat tgtctgagta ggtgtcattc
     4081 tattctgggg ggtggggtgg ggcaggacag caagggggag gattgggaag acaatagcag
     4141 gcatgctggg gacgcggccg caatttaacg cgaattaatt ctgtggaatg tgtgtcagtt
     4201 agggtgtgga aagtccccag gctccccagg caggcagaag tatgcaaagc atgcatctca
     4261 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     4321 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     4381 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     4441 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     4501 gaggcctagg cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc
     4561 aagagacagg atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc
     4621 cggccgcttg ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct
     4681 ctgatgccgc cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg
     4741 acctgtccgg tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca
     4801 cgacgggcgt tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc
     4861 tgctattggg cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga
     4921 aagtatccat catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc
     4981 cattcgacca ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc
     5041 ttgtcgatca ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg
     5101 ccaggctcaa ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct
     5161 gcttgccgaa tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc
     5221 tgggtgtggc ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc
     5281 ttggcggcga atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc
     5341 agcgcatcgc cttctatcgc cttcttgacg agttcttctg agtcgacaat caacctctgg
     5401 attacaaaat ttgtgaaaga ttgactggta ttcttaacta tgttgctcct tttacgctat
     5461 gtggatacgc tgctttaatg cctttgtatc atgcgttaac taaacttgtt tattgcagct
     5521 tataatggtt acaaataaag caatagcatc acaaatttca caaataaagc atttttttca
     5581 ctgcattcta gttgtggttt gtccaaactc atcaatgtat cttatcatgt ctggaattga
     5641 ctcaaatgat gtcaattagt ctatcagaag ctcatctggt ctcccttccg ggggacaaga
     5701 catccctgtt taatatttaa acagcagtgt tcccaaactg ggttcttata tcccttgctc
     5761 tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta aagaaacagt
     5821 ggcagccagg tttagccccg gaattgactg gattcctttt ttagggccca ttggtatggc
     5881 tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa gcgttcagag
     5941 gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct gccggctcgg
     6001 ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc tgccccctag
     6061 cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc ctgatatcta
     6121 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     6181 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     6241 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     6301 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     6361 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
     6421 gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc
     6481 aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga
     6541 agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg
     6601 cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt
     6661 cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt
     6721 cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg
     6781 tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc
     6841 tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg
     6901 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     6961 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     7021 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     7081 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     7141 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     7201 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     7261 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     7321 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     7381 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     7441 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     7501 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     7561 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     7621 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     7681 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     7741 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     7801 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     7861 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     7921 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     7981 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     8041 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     8101 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     8161 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     8221 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     8281 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     8341 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     8401 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     8461 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     8521 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     8581 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     8641 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     8701 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     8761 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     8821 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     8881 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact cat
//
