LOCUS       62056_8895        6375 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 6375)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..6375
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        123..1301
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     CDS             1330..2073
                     /codon_start=1
                     /label=tTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
                     /translation="MSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWHV
                     KNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGTR
                     PTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPTT
                     DSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGGPADALDDFDLDMLPA
                     DALDDFDLDMLPADALDDFDLDMLPG"
     polyA_signal    2130..2354
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        2406..2720
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     polyA_signal    3216..3337
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      complement(3344..3799)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        3826..3930
                     /label=AmpR promoter
     promoter        3932..4289
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4324..5115
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKASMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5350..5397
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      5726..6314
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cgcgttgaca ttgattattg actaggcttt tgcaaaaagc tttgcaaaga tggataaagt
       61 tttaaacaga gaggaatctt tgcagctaat ggaccttcta ggtcttgaaa ggagtgggaa
      121 ttggctccgg tgcccgtcag tgggcagagc gcacatcgcc cacagtcccc gagaagttgg
      181 ggggaggggt cggcaattga accggtgcct agagaaggtg gcgcggggta aactgggaaa
      241 gtgatgtcgt gtactggctc cgcctttttc ccgagggtgg gggagaaccg tatataagtg
      301 cagtagtcgc cgtgaacgtt ctttttcgca acgggtttgc cgccagaaca caggtaagtg
      361 ccgtgtgtgg ttcccgcggg cctggcctct ttacgggtta tggcccttgc gtgccttgaa
      421 ttacttccac ctggctgcag tacgtgattc ttgatcccga gcttcgggtt ggaagtgggt
      481 gggagagttc gaggccttgc gcttaaggag ccccttcgcc tcgtgcttga gttgaggcct
      541 ggcctgggcg ctggggccgc cgcgtgcgaa tctggtggca ccttcgcgcc tgtctcgctg
      601 ctttcgataa gtctctagcc atttaaaatt tttgatgacc tgctgcgacg ctttttttct
      661 ggcaagatag tcttgtaaat gcgggccaag atctgcacac tggtatttcg gtttttgggg
      721 ccgcgggcgg cgacggggcc cgtgcgtccc agcgcacatg ttcggcgagg cggggcctgc
      781 gagcgcggcc accgagaatc ggacgggggt agtctcaagc tggccggcct gctctggtgc
      841 ctggcctcgc gccgccgtgt atcgccccgc cctgggcggc aaggctggcc cggtcggcac
      901 cagttgcgtg agcggaaaga tggccgcttc ccggccctgc tgcagggagc tcaaaatgga
      961 ggacgcggcg ctcgggagag cgggcgggtg agtcacccac acaaaggaaa agggcctttc
     1021 cgtcctcagc cgtcgcttca tgtgactcca cggagtaccg ggcgccgtcc aggcacctcg
     1081 attagttctc gagcttttgg agtacgtcgt ctttaggttg gggggagggg ttttatgcga
     1141 tggagtttcc ccacactgag tgggtggaga ctgaagttag gccagcttgg cacttgatgt
     1201 aattctcctt ggaatttgcc ctttttgagt ttggatcttg gttcattctc aagcctcaga
     1261 cagtggttca aagttttttt cttccatttc aggtgtcgtg aggaattagc ttggtacgct
     1321 agcgccacca tgtctagact ggacaagagc aaagtcataa actctgctct ggaattactc
     1381 aatgaagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     1441 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctggcaatc
     1501 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     1561 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     1621 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     1681 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     1741 ggccacttta cactgggctg cgtattggag gatcaggagc atcaagtagc aaaagaggaa
     1801 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     1861 gaccatcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     1921 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     1981 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     2041 cttgacgatt ttgaccttga catgctcccc gggtaactag aatagaggcc ttgacattgc
     2101 tagcgtttaa acccgctgat cagcctcgac tgtgccttct agttgccagc catctgttgt
     2161 ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc actcccactg tcctttccta
     2221 ataaaatgag gaaattgcat cgcattgtct gagtaggtgt cattctattc tggggggtgg
     2281 ggtggggcag gacagcaagg gggaggattg ggaagacaat agcaggcatg ctggggatgc
     2341 ggtgggctct atggcttctg acagcagaga tccagtttat cgatgaggcc ctttcgtctt
     2401 cactcgagtt tactccctat cagtgataga gaacgtatgt cgagtttact ccctatcagt
     2461 gatagagaac gatgtcgagt ttactcccta tcagtgatag agaacgtatg tcgagtttac
     2521 tccctatcag tgatagagaa cgtatgtcga gtttactccc tatcagtgat agagaacgta
     2581 tgtcgagttt atccctatca gtgatagaga acgtatgtcg agtttactcc ctatcagtga
     2641 tagagaacgt atgtcgaggt aggcgtgtac ggtgggaggc ctatataagc agagctcgtt
     2701 tagtgaaccg tcagatcgcc tggagaagga tgagacccaa gctggctagc gaggcttcag
     2761 tactttacag aatcgttgcc tgcacatctt ggaaacactt gctgggatta cttcttcagg
     2821 ttaacccaac agaaggctaa agaaggtata ttgctgttga cagtgagcgc actctttcta
     2881 ggaggttgtg atagtgaagc cacagatgta tcacaacctc ctagaaagag tatgcctact
     2941 gcctcggact tcaaggggct actttaggag caattatctt gtttactaaa actgaatacc
     3001 ttgctatctc tttgatacat ttttacaaag ctgaattaaa atggtataaa tctcgagtct
     3061 agagggcccg tttaaacccg ctgatcagcc tcgagcggcc gcgactctag atcataatca
     3121 gccataccac atttgtagag gttttacttg ctttaaaaaa cctcccacac ctccccctga
     3181 acctgaaaca taaaatgaat gcaattgttg ttgttaactt gtttattgca gcttataatg
     3241 gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt tcactgcatt
     3301 ctagttgtgg tttgtccaaa ctcatcaatg tatcttaagg cgtaaattgt aagcgttaat
     3361 attttgttaa aattcgcgtt aaatttttgt taaatcagct cattttttaa ccaataggcc
     3421 gaaatcggca aaatccctta taaatcaaaa gaatagaccg agatagggtt gagtgttgtt
     3481 ccagtttgga acaagagtcc actattaaag aacgtggact ccaacgtcaa agggcgaaaa
     3541 accgtctatc agggcgatgg cccactacgt gaaccatcac cctaatcaag ttttttgggg
     3601 tcgaggtgcc gtaaagcact aaatcggaac cctaaaggga gcccccgatt tagagcttga
     3661 cggggaaagc cggcgaacgt ggcgagaaag gaagggaaga aagcgaaagg agcgggcgct
     3721 agggcgctgg caagtgtagc ggtcacgctg cgcgtaacca ccacacccgc cgcgcttaat
     3781 gcgccgctac agggcgcgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3841 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3901 atgcttcaat aatattgaaa aaggaagagt cctgaggcgg aaagaaccag ctgtggaatg
     3961 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     4021 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     4081 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     4141 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     4201 ttatttatgc agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag
     4261 gcttttttgg aggcctaggc ttttgcaaag atcgatcaag agacaggatg aggatcgttt
     4321 cgcatgattg aacaagatgg attgcacgca ggttctccgg ccgcttgggt ggagaggcta
     4381 ttcggctatg actgggcaca acagacaatc ggctgctctg atgccgccgt gttccggctg
     4441 tcagcgcagg ggcgcccggt tctttttgtc aagaccgacc tgtccggtgc cctgaatgaa
     4501 ctgcaagacg aggcagcgcg gctatcgtgg ctggccacga cgggcgttcc ttgcgcagct
     4561 gtgctcgacg ttgtcactga agcgggaagg gactggctgc tattgggcga agtgccgggg
     4621 caggatctcc tgtcatctca ccttgctcct gccgagaaag tatccatcat ggctgatgca
     4681 atgcggcggc tgcatacgct tgatccggct acctgcccat tcgaccacca agcgaaacat
     4741 cgcatcgagc gagcacgtac tcggatggaa gccggtcttg tcgatcagga tgatctggac
     4801 gaagagcatc aggggctcgc gccagccgaa ctgttcgcca ggctcaaggc gagcatgccc
     4861 gacggcgagg atctcgtcgt gacccatggc gatgcctgct tgccgaatat catggtggaa
     4921 aatggccgct tttctggatt catcgactgt ggccggctgg gtgtggcgga ccgctatcag
     4981 gacatagcgt tggctacccg tgatattgct gaagagcttg gcggcgaatg ggctgaccgc
     5041 ttcctcgtgc tttacggtat cgccgctccc gattcgcagc gcatcgcctt ctatcgcctt
     5101 cttgacgagt tcttctgagc gggactctgg ggttcgaaat gaccgaccaa gcgacgccca
     5161 acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg ggcttcggaa
     5221 tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg ctggagttct
     5281 tcgcccaccc tagggggagg ctaactgaaa cacggaagga gacaataccg gaaggaaccc
     5341 gcgctatgac ggcaataaaa agacagaata aaacgcacgg tgttgggtcg tttgttcata
     5401 aacgcggggt tcggtcccag ggctggcact ctgtcgatac cccaccgaga ccccattggg
     5461 gccaatacgc ccgcgtttct tccttttccc caccccaccc cccaagttcg ggtgaaggcc
     5521 cagggctcgc agccaacgtc ggggcggcag gccctgccat agcctcaggt tactcatata
     5581 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     5641 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     5701 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     5761 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     5821 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag
     5881 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     5941 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     6001 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     6061 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat
     6121 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     6181 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     6241 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     6301 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     6361 cttttgctca catgt
//
