LOCUS       62056_12825        5753 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 5753)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..5753
                     /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"
     polyA_signal    2814..2935
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3732..3748)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(3756..3772)
                     /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(3780..3810)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(3825..3846)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4134..4722)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(4896..5753)
                     /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 ggcccgcggc cgccccggga cgcgtcatat
     2761 ggaattctac ctacgctatg tggatacgct gctttaatgc ctttgtatca gttaacttgt
     2821 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     2881 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttaatcta
     2941 taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa taacaatata
     3001 attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt cattttgact
     3061 cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc acgcctcacg
     3121 ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg cgctatttag
     3181 aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta tctttctagg
     3241 gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca gtcatcgccc
     3301 aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc gcgaggttga
     3361 agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg agcgaaaacg
     3421 cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg gtgaactgtt
     3481 cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt tccggatggt
     3541 cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact gccgtgccgg
     3601 tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg acgggtatcc
     3661 tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg ggcgcgcttg
     3721 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     3781 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     3841 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     3901 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     3961 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     4021 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     4081 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     4141 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     4201 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     4261 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     4321 ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     4381 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     4441 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     4501 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     4561 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     4621 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     4681 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     4741 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     4801 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     4861 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     4921 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     4981 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     5041 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     5101 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     5161 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     5221 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     5281 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     5341 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     5401 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     5461 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     5521 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     5581 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     5641 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     5701 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact cat
//
