LOCUS       62056_12820        7863 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 7863)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7863
                     /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"
     promoter        1449..1816
                     /label=TRE3GS promoter
                     /note="3rd-generation Tet-responsive promoter that can be 
                     activated by binding of Tet-On(R) 3G, modified to eliminate
                     binding sites for endogenous mammalian transcription 
                     factors"
     promoter        1883..2393
                     /label=hPGK promoter
                     /note="human phosphoglycerate kinase 1 promoter"
     CDS             2412..3155
                     /codon_start=1
                     /label=Tet-On(R) 3G
                     /note="modified rtTA protein that binds tightly to
                     promoters containing the tet operator in the presence of 
                     doxycycline"
                     /translation="MSRLDKSKVINSALELLNGVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALPIEMLDRHHTHSCPLEGESWQDFLRNNAKSYRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEEQEHQVAKEERETPTT
                     DSMPPLLKQAIELFDRQGAEPAFLFGLELIICGLEKQLKCESGGPTDALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     promoter        3200..3411
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3424..3692
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3727..4323
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4434..4566
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5842..5858)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5866..5882)
                     /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(5890..5920)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5935..5956)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6244..6832)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7006..7863)
                     /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 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 gtattatgcc cagtacatga
     1381 ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccaatt
     1441 atcacctcga gtttactccc tatcagtgat agagaacgta tgaagagttt actccctatc
     1501 agtgatagag aacgtatgca gactttactc cctatcagtg atagagaacg tataaggagt
     1561 ttactcccta tcagtgatag agaacgtatg accagtttac tccctatcag tgatagagaa
     1621 cgtatctaca gtttactccc tatcagtgat agagaacgta tatccagttt actccctatc
     1681 agtgatagag aacgtataag ctttgcttat gtaaaccagg gcgcctataa aagagtgctg
     1741 attttttgag taaacttcaa ttccacaaca cttttgtctt ataccaactt tccgtaccac
     1801 ttcctaccct cgtaaagaat tcccgtatac accggttgct ggacttcgat tcgagcggat
     1861 ccactcgagg tgataattcc acggggttgg ggttgcgcct tttccaaggc agccctgggt
     1921 ttgcgcaggg acgcggctgc tctgggcgtg gttccgggaa acgcagcggc gccgaccctg
     1981 ggtctcgcac attcttcacg tccgttcgca gcgtcacccg gatcttcgcc gctacccttg
     2041 tgggcccccc ggcgacgctt cctgctccgc ccctaagtcg ggaaggttcc ttgcggttcg
     2101 cggcgtgccg gacgtgacaa acggaagccg cacgtctcac tagtaccctc gcagacggac
     2161 agcgccaggg agcaatggca gcgcgccgac cgcgatgggc tgtggccaat agcggctgct
     2221 cagcagggcg cgccgagagc agcggccggg aaggggcggt gcgggaggcg gggtgtgggg
     2281 cggtagtgtg ggccctgttc ctgcccgcgc ggtgttccgc attctgcaag cctccggagc
     2341 gcacgtcggc agtcggctcc ctcgttgacc gaatcaccga cctctctccc cagggggatc
     2401 atcgaattac catgtctaga ctggacaaga gcaaagtcat aaactctgct ctggaattac
     2461 tcaatggagt cggtatcgaa ggcctgacga caaggaaact cgctcaaaag ctgggagttg
     2521 agcagcctac cctgtactgg cacgtgaaga acaagcgggc cctgctcgat gccctgccaa
     2581 tcgagatgct ggacaggcat catacccact cctgccccct ggaaggcgag tcatggcaag
     2641 actttctgcg gaacaacgcc aagtcatacc gctgtgctct cctctcacat cgcgacgggg
     2701 ctaaagtgca tctcggcacc cgcccaacag agaaacagta cgaaaccctg gaaaatcagc
     2761 tcgcgttcct gtgtcagcaa ggcttctccc tggagaacgc actgtacgct ctgtccgccg
     2821 tgggccactt tacactgggc tgcgtattgg aggaacagga gcatcaagta gcaaaagagg
     2881 aaagagagac acctaccacc gattctatgc ccccacttct gaaacaagca attgagctgt
     2941 tcgaccggca gggagccgaa cctgccttcc ttttcggcct ggaactaatc atatgtggcc
     3001 tggagaaaca gctaaagtgc gaaagcggcg ggccgaccga cgcccttgac gattttgact
     3061 tagacatgct cccagccgat gcccttgacg actttgacct tgatatgctg cctgctgacg
     3121 ctcttgacga ttttgacctt gacatgctcc ccgggtaagc ggccgcgaag gatctgcgat
     3181 cgctccggtg cccgtcagtg ggcagagcgc acatcgccca cagtccccga gaagttgggg
     3241 ggaggggtcg gcaattgaac gggtgcctag agaaggtggc gcggggtaaa ctgggaaagt
     3301 gatgtcgtgt actggctccg cctttttccc gagggtgggg gagaaccgta tataagtgca
     3361 gtagtcgccg tgaacgttct ttttcgcaac gggtttgccg ccagaacaca gctgaagctt
     3421 cgaggggctc gcatctctcc ttcacgcgcc cgccgcccta cctgaggccg ccatccacgc
     3481 cggttgagtc gcgttctgcc gcctcccgcc tgtggtgcct cctgaactgc gtccgccgtc
     3541 taggtaagtt taaagctcag gtcgagaccg ggcctttgtc cggcgctccc ttggagccta
     3601 cctagactca gccggctctc cacgctttgc ctgaccctgc ttgctcaact ctacgtcttt
     3661 gtttcgtttt ctgttctgcg ccgttacaga tccaagctgt gaccggcgcc tacgctagac
     3721 gccaccatga ccgagtacaa gcccacggtg cgcctcgcca cccgcgacga cgtccccagg
     3781 gccgtacgca ccctcgccgc cgcgttcgcc gactaccccg ccacgcgcca caccgtcgat
     3841 ccggaccgcc acatcgagcg ggtcaccgag ctgcaagaac tcttcctcac gcgcgtcggg
     3901 ctcgacatcg gcaaggtgtg ggtcgcggac gacggcgccg cggtggcggt ctggaccacg
     3961 ccggagagcg tcgaagcggg ggcggtgttc gccgagatcg gcccgcgcat ggccgagttg
     4021 agcggttccc ggctggccgc gcagcaacag atggaaggcc tcctggcgcc gcaccggccc
     4081 aaggagcccg cgtggttcct ggccaccgtc ggcgtctcgc ccgaccacca gggcaagggt
     4141 ctgggcagcg ccgtcgtgct ccccggagtg gaggcggccg agcgcgccgg ggtgcccgcc
     4201 ttcctggaga cctccgcgcc ccgcaacctc cccttctacg agcggctcgg cttcaccgtc
     4261 accgccgacg tcgaggtgcc cgaaggaccg cgcacctggt gcatgacccg caagcccggt
     4321 gcctgaaatc aacctctgga ttacaaaatt tgtgaaagat tgactggtat tcttaactat
     4381 gttgctcctt ttacgctatg tggatacgct gctttaatgc ctttgtatca gttaacttgt
     4441 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     4501 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     4561 tctggaattg actcaaatga tgtcaattag tctatcagaa gctatctggt ctcccttccg
     4621 ggggacaaga catccctgtt taatatttaa acagcagtgt tcccaaactg ggttcttata
     4681 tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc tcacaggaac gaagtcccta
     4741 aagaaacagt ggcagccagg tttagccccg gaattgactg gattcctttt ttagggccca
     4801 ttggtatggc tttttccccg tatcccccca ggtgtctgca ggctcaaaga gcagcgagaa
     4861 gcgttcagag gaaagcgatc ccgtgccacc ttccccgtgc ccgggctgtc cccgcacgct
     4921 gccggctcgg ggatgcgggg ggagcgccgg accggagcgg agccccgggc ggctcgctgc
     4981 tgccccctag cgggggaggg acgtaattac atccctgggg gctttggggg ggggctgtcc
     5041 ctgatatcta taacaagaaa atatatatat aataagttat cacgtaagta gaacatgaaa
     5101 taacaatata attatcgtat gagttaaatc ttaaaagtca cgtaaaagat aatcatgcgt
     5161 cattttgact cacgcggtcg ttatagttca aaatcagtga cacttaccgc attgacaagc
     5221 acgcctcacg ggagctccaa gcggcgactg agatgtccta aatgcacagc gacggattcg
     5281 cgctatttag aaagagagag caatatttca agaatgcatg cgtcaatttt acgcagacta
     5341 tctttctagg gttaatctag ctgcatcagg atcatatcgt cgggtctttt ttccggctca
     5401 gtcatcgccc aagctggcgc tatctgggca tcggggagga agaagcccgt gccttttccc
     5461 gcgaggttga agcggcatgg aaagagtttg ccgaggatga ctgctgctgc attgacgttg
     5521 agcgaaaacg cacgtttacc atgatgattc gggaaggtgt ggccatgcac gcctttaacg
     5581 gtgaactgtt cgttcaggcc acctgggata ccagttcgtc gcggcttttc cggacacagt
     5641 tccggatggt cagcccgaag cgcatcagca acccgaacaa taccggcgac agccggaact
     5701 gccgtgccgg tgtgcagatt aatgacagcg gtgcggcgct gggatattac gtcagcgagg
     5761 acgggtatcc tggctggatg ccgcagaaat ggacatggat accccgtgag ttacccggcg
     5821 ggcgcgcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac
     5881 aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt
     5941 gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc
     6001 gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg
     6061 ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt
     6121 atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa
     6181 gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc
     6241 gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag
     6301 gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt
     6361 gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg
     6421 aagcgtggcg ctttctcata gctcacgctg taggtatctc agttcggtgt aggtcgttcg
     6481 ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg
     6541 taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac
     6601 tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg
     6661 gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt
     6721 taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg
     6781 tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc
     6841 tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt
     6901 ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt
     6961 taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag
     7021 tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt
     7081 cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg caatgatacc
     7141 gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc
     7201 cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg
     7261 ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac
     7321 aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg
     7381 atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc
     7441 tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact
     7501 gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc
     7561 aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat
     7621 acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc
     7681 ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac
     7741 tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa
     7801 aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact
     7861 cat
//
