LOCUS       62056_21435        8459 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 8459)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8459
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     polyA_signal    complement(1365..1446)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(1575..2291)
                     /codon_start=1
                     /label=EBFP2
                     /note="enhanced blue variant of GFP (Ai et al., 2007)"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVRGEGEGDATNGKLTL
                     KFICTTGKLPVPWPTLVTTLSHGVQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GTYKTRAEVKFEGDTLVNRIELKGVDFKEDGNILGHKLEYNFNSHNIYIMAVKQKNGIK
                     VNFKIRHNVEDGSVQLADHYQQNTPIGDGPVLLPDSHYLSTQSVLSKDPNEKRDHMVLL
                     EFRTAAGITLGMDELYK"
     promoter        complement(2330..2348)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     promoter        2374..2723
                     /label=TRE3GV promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, optimized for 
                     retroviral and lentiviral vectors"
     CDS             3054..3110
                     /codon_start=1
                     /label=P2A
                     /note="2A peptide from porcine teschovirus-1 polyprotein"
                     /translation="ATNFSLLKQAGDVEENPGP"
     polyA_signal    5520..5641
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6438..6454)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6462..6478)
                     /label=lac operator
                     /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)."
     promoter        complement(6486..6516)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6531..6552)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6840..7428)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7602..8459)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
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 ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtgcagtg aaaaaaatgc
     1381 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     1441 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     1501 gttttttaaa gcaagtaaaa cctctacaaa tgtggtatgg ctgattatga tcctctagac
     1561 atatgctgca gtcacttgta cagctcatcc atgcccaggg tgatgccagc ggcggtccga
     1621 aattccagca gcaccatgtg gtcccgcttc tcgttggggt ccttgctcag cacgctctgg
     1681 gtgctcaggt agtggctatc aggcagcagc acggggccat ctccgatggg ggtgttctgc
     1741 tggtagtggt cggccagctg cacgctgcca tcttccacgt tgtgccggat cttgaagttc
     1801 actttgatgc cgtttttctg cttcacggcc atgatgtaga tgttgtggct gttgaagttg
     1861 tactccagct tgtggcccag gatgttgccg tcctctttga agtccacgcc cttcagctcg
     1921 atccggttca cgagggtgtc gccctcgaac ttcacttcgg ctctggtctt gtaggtgccg
     1981 tcgtccttga agaagatggt ccgttcctgc acgtagccct cgggcatggc gctcttgaag
     2041 aaatcgtgct gcttcatgtg gtcggggtat ctggcgaagc actgcacgcc gtgagacagt
     2101 gtggtcacga gggtaggcca aggcacgggc agcttgccgg tggtgcagat gaacttcagg
     2161 gtcagcttgc catttgtggc gtcgccttcg ccctctcccc gcacagagaa cttgtggccg
     2221 ttcacgtcgc catccagttc caccaggatg ggcaccacgc cggtgaacag ttcctcgccc
     2281 ttggacacca tggtgaaggg tactgagctc ggtacctgca ggcgtacctt ctatagtgtc
     2341 acctaaatgc gatctgacgg ttcacttata ctcgagttta ctccctatca gtgatagaga
     2401 acgtatgaag agtttactcc ctatcagtga tagagaacgt atgcagactt tactccctat
     2461 cagtgataga gaacgtataa ggagtttact ccctatcagt gatagagaac gtatgaccag
     2521 tttactccct atcagtgata gagaacgtat ctacagttta ctccctatca gtgatagaga
     2581 acgtatatcc agtttactcc ctatcagtga tagagaacgt atgtcgaggt aggcgtgtac
     2641 ggtgggcgcc tataaaagca gagctcgttt agtgaaccgt cagatcgcct ggagcaattc
     2701 cacaacactt ttgtcttata cttggatccg ccaccatggc cgtggaacac aacatgcaga
     2761 gcagcaccat cgtggacgtg ctggaaaaga tcctggacaa gggcgtcgtg atcgccgggg
     2821 acatcacagt gggaatcgcc gacgtggaac tgctgaccat caagatccgg ctgatcgtgg
     2881 ccagcgtgga caaggccaaa gaaatcggca tggattggtg ggagaacgac ccctacctga
     2941 gcagcaaggg cgccaacaac aaggctctgg aagaggaaaa caagatgctg cacgagcggc
     3001 tgaaaacact ggaagagaag atcgagacaa agcgctatag aagcgccatt ggcgcaacaa
     3061 attttagtct tttaaagcag gcaggagacg tcgaggaaaa ccctggaccc gtgatgagtg
     3121 aaacaaacga aacaggtatt tatattttta gcgccattca aacggataaa gacgaagaat
     3181 ttggcgccgt ggaagtagaa ggaacaaaag ctgaaacatt tttgattcgc tacaaagacg
     3241 cggctatggt agcagctgaa gtaccgatga aaatttatca tcctaatcgc caaaatttat
     3301 taatgcatca aaacgcagta gcagcgatta tggacaagaa cgatacggtt attccaatca
     3361 gctttgggaa tgtattcaaa tcaaaagaag acgtaaaagt tcttttggaa aacctttatc
     3421 cgcagtttga aaagctgttt ccagcgatca aaggaaaaat tgaagtcggt ttaaaagtaa
     3481 ttgggaaaaa agaatggctt gagaaaaaag taaacgaaaa tcctgaactt gagaaagtat
     3541 cagcatccgt aaaaggaaaa tcagaagcag ccggttatta tgagcgtatt caacttggag
     3601 gaatggctca aaagatgttt acttccctgc aaaaagaagt caagacagat gtgttttctc
     3661 cgcttgaaga agcagcggaa gcagcaaaag caaatgagcc aacgggcgaa acgatgcttt
     3721 taaacgcgtc tttcttaatt aaccgagaag atgaagcgaa gtttgatgaa aaagtaaatg
     3781 aagcgcatga aaactggaaa gacaaagccg attttcatta cagcggtcct tggcctgctt
     3841 ataattttgt gaacattcgc ctaaaagtag aagagaaagc acctggctcg ggagcgacca
     3901 acttctcatt actcaaacaa gccggagacg ttgaggagaa tccaggccct gtgctgcaca
     3961 agctcgtgac cgcccccatc aacctggtcg tgaagatcgg cgagaaggtg caggaagagg
     4021 ccgacaagca gctgtacgac ctgcccacca tccagcagaa gctgatccag ctgcagatga
     4081 tgttcgagct gggcgagatc cccgaggaag ccttccagga aaaagaggac gaactgctga
     4141 tgagatacga gatcgccaag cggcgcgaga ttgagcagtg ggaagaactg acccagaagc
     4201 ggaatgagga aagcaacgca gagaaattga acgccactaa cttctccctg ttgaaacaag
     4261 caggggatgt cgaagagaat cccgggccac cgatgggaga attactgtat ttatacggtt
     4321 taattccaac aaaagaagca gcagccatag agccgtttcc atcttataag gggtttgacg
     4381 gagaacattc actgtaccca attgcgtttg atcaggtgac ggctgtagtt tctaagctgg
     4441 atgctgacac ctattcagaa aaagtgattc aagaaaaaat ggagcaggat atgagctggc
     4501 tgcaggaaaa agcatttcat catcacgaaa cggtagccgc tttgtacgaa gaatttacga
     4561 tcattccatt aaaattttgc accatttata aaggtgaaga aagtctgcag gcagctattg
     4621 agattaacaa agaaaagata gagaattcac tgacgctgct tcaaggaaat gaagagtgga
     4681 atgtgaaaat ttactgtgat gatacagagc ttaaaaaagg aatcagcgaa acgaatgaaa
     4741 gcgtgaaagc gaaaaaacaa gaaattagtc acttatcacc aggaagacag ttttttgaaa
     4801 agaaaaaaat agatcagctg attgaaaaag aattagagct tcacaaaaac aaagtgtgtg
     4861 aagagataca tgacaagcta aaagaattat cgctttatga ctctgttaaa aagaattgga
     4921 gcaaagacgt aactggcgca gctgaacaga tggcgtggaa cagcgtgttt cttctcccgt
     4981 ctctgcagat tactaagttc gtaaacgaaa tagaagagct tcagcaaagg cttgaaaata
     5041 aaggctggaa gtttgaagtg acgggaccat ggccgcccta tcatttctcg agctttgcgg
     5101 gagcacctgg tagcggcgcc acgaattttt cgttgttgaa gcaagctggg gatgttgaag
     5161 agaacccagg gcctgtgcag cccgtgtccc aggccaacgg cagaatccac ctggatcccg
     5221 atcaggccga acagggactg gcccagctcg tgatgaccgt gatcgagctg ctgcggcaga
     5281 tcgtggaacg gcacgccatg agaagagtgg aaggcggcac cctgaccgac gagcagatcg
     5341 agaatctggg aatcgccctg atgaacctgg aagagaagat ggacgagctg aaagaggtgt
     5401 tcggactgga cgccgaggac ctgaacatcg acctgggccc tctgggcagc ctgctgtgag
     5461 aattctacct acgctatgtg gatacgctgc tttaatgcct ttgtatcatg cgttaactaa
     5521 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     5581 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     5641 aatctataac aagaaaatat atatataata agttatcacg taagtagaac atgaaataac
     5701 aatataatta tcgtatgagt taaatcttaa aagtcacgta aaagataatc atgcgtcatt
     5761 ttgactcacg cggtcgttat agttcaaaat cagtgacact taccgcattg acaagcacgc
     5821 ctcacgggag ctccaagcgg cgactgagat gtcctaaatg cacagcgacg gattcgcgct
     5881 atttagaaag agagagcaat atttcaagaa tgcatgcgtc aattttacgc agactatctt
     5941 tctagggtta atctagctgc atcaggatca tatcgtcggg tcttttttcc ggctcagtca
     6001 tcgcccaagc tggcgctatc tgggcatcgg ggaggaagaa gcccgtgcct tttcccgcga
     6061 ggttgaagcg gcatggaaag agtttgccga ggatgactgc tgctgcattg acgttgagcg
     6121 aaaacgcacg tttaccatga tgattcggga aggtgtggcc atgcacgcct ttaacggtga
     6181 actgttcgtt caggccacct gggataccag ttcgtcgcgg cttttccgga cacagttccg
     6241 gatggtcagc ccgaagcgca tcagcaaccc gaacaatacc ggcgacagcc ggaactgccg
     6301 tgccggtgtg cagattaatg acagcggtgc ggcgctggga tattacgtca gcgaggacgg
     6361 gtatcctggc tggatgccgc agaaatggac atggataccc cgtgagttac ccggcgggcg
     6421 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     6481 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     6541 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     6601 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     6661 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     6721 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     6781 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     6841 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     6901 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     6961 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     7021 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     7081 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     7141 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     7201 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     7261 aactacggct acactagaag gacagtattt ggtatctgcg ctctgctgaa gccagttacc
     7321 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     7381 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     7441 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     7501 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     7561 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     7621 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     7681 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     7741 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     7801 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     7861 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     7921 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     7981 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     8041 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     8101 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     8161 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     8221 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     8281 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     8341 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     8401 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcat
//
