LOCUS       62056_5195        4979 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4979)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..4979
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(268..295)
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB
                     gene"
     terminator      complement(387..473)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     primer_bind     537..553
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    569..668
                     /label=attL1
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        733..936
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     CDS             1036..1107
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags"
                     /translation="DYKDDDDKDYKDDDDKDYKDDDDK"
     promoter        1152..1363
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1376..1644
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             1700..2365
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     misc_feature    2444..3032
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     protein_bind    complement(3104..3203)
                     /label=attL2
                     /note="recombination site for the Gateway(R) LR reaction"
     promoter        complement(3221..3239)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(3244..3260)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             3373..4179
                     /codon_start=1
                     /label=KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      4329..4917
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
       61 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
      121 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
      181 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaata cgcgtaccgc
      241 tagccaggaa gagtttgtag aaacgcaaaa aggccatccg tcaggatggc cttctgctta
      301 gtttgatgcc tggcagttta tggcgggcgt cctgcccgcc accctccggg ccgttgcttc
      361 acaacgttca aatccgctcc cggcggattt gtcctactca ggagagcgtt caccgacaaa
      421 caacagataa aacgaaaggc ccagtcttcc gactgagcct ttcgttttat ttgatgcctg
      481 gcagttccct actctcgcgt taacgctagc atggatgttt tcccagtcac gacgttgtaa
      541 aacgacggcc agtcttaagc tcgggcccca aataatgatt ttattttgac tgatagtgac
      601 ctgttcgttg caacaaattg atgagcaatg cttttttata atgccaactt tgtacaaaaa
      661 agcaggctcc gcggcttatg ggactttcct acttggcagt acatctacgt attagtcatc
      721 gctattacca tggtgatgcg gttttggcag tacatcaatg ggcgtggata gcggtttgac
      781 tcacggggat ttccaagtct ccaccccatt gacgtcaatg ggagtttgtt ttggcaccaa
      841 aatcaacggg actttccaaa atgtcgtaac aactccgccc cattgacgca aatgggcggt
      901 aggcgtgtac ggtgggaggt ctatataagc agagctcgtt tagtgaaccg tcagatcgcc
      961 tggagacgcc atccacgctg ttttgacctc catagaagat tctagagcta gcgaattcga
     1021 atttaaatcg gatccgacta caaggacgac gatgacaagg actacaagga cgacgatgac
     1081 aaggactaca aggacgacga tgacaagtaa gcggccgcga aggatctgcg atcgctccgg
     1141 tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg ggggaggggt
     1201 cggcaattga acgggtgcct agagaaggtg gcgcggggta aactgggaaa gtgatgtcgt
     1261 gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg cagtagtcgc
     1321 cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca cagctgaagc ttcgaggggc
     1381 tcgcatctct ccttcacgcg cccgccgccc tacctgaggc cgccatccac gccggttgag
     1441 tcgcgttctg ccgcctcccg cctgtggtgc ctcctgaact gcgtccgccg tctaggtaag
     1501 tttaaagctc aggtcgagac cgggcctttg tccggcgctc ccttggagcc tacctagact
     1561 cagccggctc tccacgcttt gcctgaccct gcttgctcaa ctctacgtct ttgtttcgtt
     1621 ttctgttctg cgccgttaca gatccaagct gtgaccggcg cctacgctag acgccaccat
     1681 ggagagcgac gagagcggcc tgcccgccat ggagatcgag tgccgcatca ccggcaccct
     1741 gaacggcgtg gagttcgagc tggtgggcgg cggagagggc acccccaagc agggccgcat
     1801 gaccaacaag atgaagagca ccaaaggcgc cctgaccttc agcccctacc tgctgagcca
     1861 cgtgatgggc tacggcttct accacttcgg cacctacccc agcggctacg agaacccctt
     1921 cctgcacgcc atcaacaacg gcggctacac caacacccgc atcgagaagt acgaggacgg
     1981 cggcgtgctg cacgtgagct tcagctaccg ctacgaggcc ggccgcgtga tcggcgactt
     2041 caaggtggtg ggcaccggct tccccgagga cagcgtgatc ttcaccgaca agatcatccg
     2101 cagcaacgcc accgtggagc acctgcaccc catgggcgat aacgtgctgg tgggcagctt
     2161 cgcccgcacc ttcagcctgc gcgacggcgg ctactacagc ttcgtggtgg acagccacat
     2221 gcacttcaag agcgccatcc accccagcat cctgcagaac gggggcccca tgttcgcctt
     2281 ccgccgcgtg gaggagctgc acagcaacac cgagctgggc atcgtggagt accagcacgc
     2341 cttcaagacc cccatcgcct tcgccagatc ccgcgctcag tcgtccaatt ctgccgtgga
     2401 cggcaccgcc ggacccggct ccaccggatc tcgctaagtc gacaatcaac ctctggatta
     2461 caaaatttgt gaaagattga ctggtattct taactatgtt gctcctttta cgctatgtgg
     2521 atacgctgct ttaatgcctt tgtatcatgc tattgcttcc cgtatggctt tcattttctc
     2581 ctccttgtat aaatcctggt tgctgtctct ttatgaggag ttgtggcccg ttgtcaggca
     2641 acgtggcgtg gtgtgcactg tgtttgctga cgcaaccccc actggttggg gcattgccac
     2701 cacctgtcag ctcctttccg ggactttcgc tttccccctc cctattgcca cggcggaact
     2761 catcgccgcc tgccttgccc gctgctggac aggggctcgg ctgttgggca ctgacaattc
     2821 cgtggtgttg tcggggaaat catcgtcctt tccttggctg ctcgcctgtg ttgccacctg
     2881 gattctgcgc gggacgtcct tctgctacgt cccttcggcc ctcaatccag cggaccttcc
     2941 ttcccgcggc ctgctgccgg ctctgcggcc tcttccgcgt cttcgccttc gccctcagac
     3001 gagtcggatc tccctttggg ccgcctcccc gcctggtacc tttaagacca atgacttaca
     3061 aggcagcccc atacaagtat tggggataag ggtgggcgcg ccgacccagc tttcttgtac
     3121 aaagttggca ttataagaaa gcattgctta tcaatttgtt gcaacgaaca ggtcactatc
     3181 agtcaaaata aaatcattat ttgccatcca gctgatatcc cctatagtga gtcgtattac
     3241 atggtcatag ctgtttcctg gcagctctgg cccgtgtctc aaaatctctg atgttacatt
     3301 gcacaagata aaaatatatc atcatgaaca ataaaactgt ctgcttacat aaacagtaat
     3361 acaaggggtg ttatgagcca tattcaacgg gaaacgtcga ggccgcgatt aaattccaac
     3421 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     3481 acaatctatc gcttgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     3541 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     3601 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     3661 actgcgatcc ccggaaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     3721 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     3781 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     3841 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     3901 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     3961 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     4021 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     4081 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     4141 aataaattgc agtttcattt gatgctcgat gagtttttct aatcagaatt ggttaattgg
     4201 ttgtaacact ggcagagcat tacgctgact tgacgggacg gcgcaagctc atgaccaaaa
     4261 tcccttaacg tgagttacgc gtcgttccac tgagcgtcag accccgtaga aaagatcaaa
     4321 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     4381 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     4441 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     4501 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     4561 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     4621 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     4681 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     4741 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     4801 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     4861 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4921 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgtt
//
