LOCUS       Exported                6454 bp ds-DNA     circular SYN 16-JAN-2017
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6454)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Monday, January 16, 2017 from SnapGene Viewer 3.1.2
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..6454
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     promoter        16..1178
                     /note="EF-1-alpha promoter"
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          247..1169
                     /note="EF-1-alpha intron A"
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     intron          1217..1349
                     /note="chimeric intron"
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
     promoter        1394..1412
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    1418..1471
                     /note="BIP signal peptide"
     CDS             1472..2167
                     /codon_start=1
                     /product="monomeric blue fluorescent protein"
                     /note="TagBFP"
                     /note="mammalian codon-optimized"
                     /translation="SELIKENMHMKLYMEGTVDNHHFKCTSEGEGKPYEGTQTMRIKVV
                     EGGPLPFAFDILATSFLYGSKTFINHTQGIPDFFKQSFPEGFTWERVTTYEDGGVLTAT
                     QDTSLQDGCLIYNVKIRGVNFTSNGPVMQKKTLGWEAFTETLYPADGGLEGRNDMALKL
                     VGGSHLIANIKTTYRSKKPAKNLKMPGVYYVDYRLERIKEANNETYVEQHEVAVARYCD
                     LPSKLGHKLN"
     misc_feature    2245..2817
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     misc_feature    2268..2819
                     /note="IRES"
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             2819..3421
                     /codon_start=1
                     /gene="pac from Streptomyces"
                     /product="puromycin N-acetyltransferase"
                     /note="PuroR"
                     /note="confers resistance to puromycin"
                     /translation="MGTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIE
                     RVTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRL
                     AAQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLET
                     SAPRNLPFYERLGFTVTADVECPKDRATWCMTRKPGA"
     polyA_signal    3655..3789
                     /note="SV40 poly(A) signal"
                     /note="SV40 polyadenylation signal"
     rep_origin      3864..4319
                     /direction=RIGHT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4653..4757
                     /gene="bla"
                     /note="AmpR promoter"
     CDS             4758..5618
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /note="AmpR"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      5789..6377
                     /direction=RIGHT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 attcaagctt cgtgaggctc cggtgcccgt cagtgggcag agcgcacatc gcccacagtc
       61 cccgagaagt tggggggagg ggtcggcaat tgaaccggtg cctagagaag gtggcgcggg
      121 gtaaactggg aaagtgatgt cgtgtactgg ctccgccttt ttcccgaggg tgggggagaa
      181 ccgtatataa gtgcagtagt cgccgtgaac gttctttttc gcaacgggtt tgccgccaga
      241 acacaggtaa gtgccgtgtg tggttcccgc gggcctggcc tctttacggg ttatggccct
      301 tgcgtgcctt gaattacttc cacctggctc cagtacgtga ttcttgatcc cgagctggag
      361 ccaggggcgg gccttgcgct ttaggagccc cttcgcctcg tgcttgagtt gaggcctggc
      421 ctgggcgctg gggccgccgc gtgcgaatct ggtggcacct tcgcgcctgt ctcgctgctt
      481 tcgataagtc tctagccatt taaaattttt gatgacctgc tgcgacgctt tttttctggc
      541 aagatagtct tgtaaatgcg ggccaggatc tgcacactgg tatttcggtt tttgggcccg
      601 cggccggcga cggggcccgt gcgtcccagc gcacatgttc ggcgaggcgg ggcctgcgag
      661 cgcggccacc gagaatcgga cgggggtagt ctcaagctgg ccggcctgct ctggtgcctg
      721 gcctcgcgcc gccgtgtatc gccccgccct gggcggcaag gctggcccgg tcggcaccag
      781 ttgcgtgagc ggaaagatgg ccgcttcccg gccctgctcc agggggctca aaatggagga
      841 cgcggcgctc gggagagcgg gcgggtgagt cacccacaca aaggaaaagg gcctttccgt
      901 cctcagccgt cgcttcatgt gactccacgg agtaccgggc gccgtccagg cacctcgatt
      961 acttctggag cttttggagt acgtcgtctt taggttgggg ggaggggttt tatgcgatgg
     1021 agtttcccca cactgagtgg gtggagactg aagttaggcc agcttggcac ttgatgtaat
     1081 tctccttgga atttggcctt tttgagtttg gatcttggtt cattctcaag cctcagacag
     1141 tggttcaaag tttttttctt ccatttcagg tgtcgtgaac acgggctgca gaagttggtc
     1201 gtgaggcact gggcaggtaa gtatcaaggt tacaagacag gtttaaggag accaatagaa
     1261 actgggcttg tcgagacaga gaagactctt gcgtttctga taggcaccta ttggtcttac
     1321 tgacatccac tttgcctttc tctccacagg tgtccactcc cagttcaatt acagctctta
     1381 aggctagagt acttaatacg actcactata ggctagcatg aagctctccc tggtggccgc
     1441 gatgctgctg ctgctcagcg cggcgcgggc cagcgagctg attaaggaga acatgcacat
     1501 gaagctgtac atggagggca ccgtggacaa ccatcacttc aagtgcacat ccgagggcga
     1561 aggcaagccc tacgagggca cccagaccat gagaatcaag gtggtcgagg gcggccctct
     1621 ccccttcgcc ttcgacatcc tggctactag cttcctctac ggcagcaaga ccttcatcaa
     1681 ccacacccag ggcatccccg acttcttcaa gcagtccttc cctgagggct tcacatggga
     1741 gagagtcacc acatacgaag acgggggcgt gctgaccgct acccaggaca ccagcctcca
     1801 ggacggctgc ctcatctaca acgtcaagat cagaggggtg aacttcacat ccaacggccc
     1861 tgtgatgcag aagaaaacac tcggctggga ggccttcacc gagacgctgt accccgctga
     1921 cggcggcctg gaaggcagaa acgacatggc cctgaagctc gtgggcggga gccatctgat
     1981 cgcaaacatc aagaccacat atagatccaa gaaacccgct aagaacctca agatgcctgg
     2041 cgtctactat gtggactaca gactggaaag aatcaaggag gccaacaacg agacctacgt
     2101 cgagcagcac gaggtggcag tggccagata ctgcgacctc cctagcaaac tggggcacaa
     2161 gcttaataaa gacgagctat gagtcgagaa ttcacgcgtg gtacctctag agtcgacccg
     2221 ggcggccgct ctagcccaat tccgcccctc tccctccccc ccccctaacg ttactggccg
     2281 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
     2341 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
     2401 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
     2461 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
     2521 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
     2581 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
     2641 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     2701 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     2761 cgtctaggcc ccccgaacca cggggacgtg gttttccttt gaaaaacacg atgataatat
     2821 gggcaccgag tacaagccca cggtgcgcct cgccacccgc gacgacgtcc cccgggccgt
     2881 acgcaccctc gccgccgcgt tcgccgacta ccccgccacg cgccacaccg tcgacccgga
     2941 ccgccacatc gagcgggtca ccgagctgca agaactcttc ctcacgcgcg tcgggctcga
     3001 catcggcaag gtgtgggtcg cggacgacgg cgccgcggtg gcggtctgga ccacgccgga
     3061 gagcgtcgaa gcgggggcgg tgttcgccga gatcggcccg cgcatggccg agttgagcgg
     3121 ttcccggctg gccgcgcagc aacagatgga aggcctcctg gcgccgcacc ggcccaagga
     3181 gcccgcgtgg ttcctggcca ccgtcggcgt ctcgcccgac caccagggca agggtctggg
     3241 cagcgccgtc gtgctccccg gagtggaggc ggccgagcgc gccggggtgc ccgccttcct
     3301 ggagacctcc gcgccccgca acctcccctt ctacgagcgg ctcggcttca ccgtcaccgc
     3361 cgacgtcgag tgcccgaagg accgcgcgac ctggtgcatg acccgcaagc ccggtgcctg
     3421 acgcccgccc cacgacccgc agcgcccgac cgaaaggagc gcacgacccc atggctccga
     3481 ccgaagccga cccgggcggc cccgccgacc ccgcacccgc ccccgaggcc caccgactct
     3541 agctagagga tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac
     3601 ctcccacacc tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg
     3661 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     3721 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     3781 gtctggatcc gggctggcgt aatagcgaag aggcccgcac cgatcgccct tcccaacagt
     3841 tgcgcagcct gaatggcgaa tggacgcgcc ctgtagcggc gcattaagcg cggcgggtgt
     3901 ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc
     3961 tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg
     4021 gctcccttta gggttccgat ttagagcttt acggcacctc gaccgcaaaa aacttgattt
     4081 gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt
     4141 ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat
     4201 ctcggtctat tcttttgatt tataagggat tttgccgatt tcggcctatt ggttaaaaaa
     4261 tgagctgatt taacaaatat ttaacgcgaa ttttaacaaa atattaacgt ttacaatttc
     4321 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
     4381 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
     4441 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
     4501 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
     4561 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
     4621 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
     4681 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
     4741 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
     4801 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
     4861 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
     4921 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
     4981 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
     5041 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
     5101 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     5161 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     5221 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     5281 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     5341 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     5401 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     5461 ccggtgagcg tgggtctcgc ggtatcattg cagcactggg gccagatggt aagccctccc
     5521 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     5581 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     5641 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     5701 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     5761 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     5821 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     5881 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgtccttc
     5941 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     6001 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     6061 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     6121 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     6181 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     6241 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     6301 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     6361 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     6421 ggccttttgc tcacatggct cgacagatcc ccca
//
