LOCUS       62056_12585        8114 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 8114)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8114
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        48..502
                     /label=TERT
                     /note="Human telomerase reverse transcriptase promoter"
     misc_feature    565..1138
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     CDS             1145..2794
                     /codon_start=1
                     /label=luciferase
                     /note="firefly luciferase"
                     /translation="MEDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGTIAFTDA
                     HIEVNITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAP
                     ANDIYNERELLNSMNISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQS
                     MYTFVTSHLPPGFNEYDFVPESFDRDKTIALIMNSSGSTGLPKGVALPHRTACVRFSHA
                     RDPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYK
                     IQSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGY
                     GLTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMS
                     GYVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESIL
                     LQHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVV
                     FVDEVPKGLTGKLDARKIREILIKAKKGGKSKL"
     CDS             2801..2866
                     /codon_start=1
                     /label=3xFLAG
                     /note="three tandem FLAG(R) epitope tags, followed by an 
                     enterokinase cleavage site"
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
     misc_feature    2890..3478
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional
                     regulatory element"
     polyA_signal    3544..3592
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal (Cole and Stacy, 1985)"
     rep_origin      3794..4222
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        4236..4565
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     CDS             4632..5423
                     /codon_start=1
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     polyA_signal    5602..5735
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5772..5788)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5796..5812)
                     /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(5820..5850)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5865..5886)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6174..6762)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6936..7793)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(7794..7898)
                     /label=AmpR promoter
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtgg cccctccctc
       61 gggttacccc acagcttagg ccgattcgac ctctctccgc tggggccctc gctggcgtcc
      121 ctgcaccctg ggagcgcgag cggcgcgcgg gcggggaagc gcggcccaga cccccgggtc
      181 cgcccggagc agctgcgctg tcggggccag gccgggctcc cagtggattc gcgggcacag
      241 acgcccagga ccgcgcttcc cacgtggcgg agggactggg gacccgggca cccgtcctgc
      301 cccttcacct tccagctccg cctcctccgc gcggaccccg ccccgtcccg acccctcccg
      361 ggtccccggc ccagccccct ccgggccctc ccagcccctc cccttccttt ccgcggcccc
      421 gccctctcct cgcggcgcga gtttcaggca gcgctgcgtc ctgctgcgca cgtgggaagc
      481 cctggccccg gccacccccg cgctagcgtt taaacttaag cttggtaccg agctcggatc
      541 cactagtcca gtgtggtgga attccccctc tccctccccc ccccctaacg ttactggccg
      601 aagccgcttg gaataaggcc ggtgtgcgtt tgtctatatg ttattttcca ccatattgcc
      661 gtcttttggc aatgtgaggg cccggaaacc tggccctgtc ttcttgacga gcattcctag
      721 gggtctttcc cctctcgcca aaggaatgca aggtctgttg aatgtcgtga aggaagcagt
      781 tcctctggaa gcttcttgaa gacaaacaac gtctgtagcg accctttgca ggcagcggaa
      841 ccccccacct ggcgacaggt gcctctgcgg ccaaaagcca cgtgtataag atacacctgc
      901 aaaggcggca caaccccagt gccacgttgt gagttggata gttgtggaaa gagtcaaatg
      961 gctctcctca agcgtattca acaaggggct gaaggatgcc cagaaggtac cccattgtat
     1021 gggatctgat ctggggcctc ggtgcacatg ctttacatgt gtttagtcga ggttaaaaaa
     1081 acgtctaggc cccccgaacc acggggacgt ggttttcctt tgaaaaacac gatgataagc
     1141 caccatggaa gacgccaaaa acataaagaa aggcccggcg ccattctatc ctcttgagga
     1201 tggaaccgct ggagagcaac tgcataaggc tatgaagaga tacgccctgg ttcctggaac
     1261 aattgctttt acagatgcac atatcgaggt gaacatcacg tacgcggaat acttcgaaat
     1321 gtccgttcgg ttggcagaag ctatgaaacg atatgggctg aatacaaatc acagaatcgt
     1381 cgtatgcagt gaaaactctc ttcaattctt tatgccggtg ttgggcgcgt tatttatcgg
     1441 agttgcagtt gcgcccgcga acgacattta taatgaacgt gaattgctca acagtatgaa
     1501 catttcgcag cctaccgtag tgtttgtttc caaaaagggg ttgcaaaaaa ttttgaacgt
     1561 gcaaaaaaaa ttaccaataa tccagaaaat tattatcatg gattctaaaa cggattacca
     1621 gggatttcag tcgatgtaca cgttcgtcac atctcatcta cctcccggtt ttaatgaata
     1681 cgattttgta ccagagtcct ttgatcgtga caaaacaatt gcactgataa tgaactcctc
     1741 tggatctact gggttaccta agggtgtggc ccttccgcat agaactgcct gcgtcagatt
     1801 ctcgcatgcc agagatccta tttttggcaa tcaaatcatt ccggatactg cgattttaag
     1861 tgttgttcca ttccatcacg gttttggaat gtttactaca ctcggatatt tgatatgtgg
     1921 atttcgagtc gtcttaatgt atagatttga agaagagctg tttttacgat cccttcagga
     1981 ttacaaaatt caaagtgcgt tgctagtacc aaccctattt tcattcttcg ccaaaagcac
     2041 tctgattgac aaatacgatt tatctaattt acacgaaatt gcttctgggg gcgcacctct
     2101 ttcgaaagaa gtcggggaag cggttgcaaa acgcttccat cttccaggga tacgacaagg
     2161 atatgggctc actgagacta catcagctat tctgattaca cccgaggggg atgataaacc
     2221 gggcgcggtc ggtaaagttg ttccattttt tgaagcgaag gttgtggatc tggataccgg
     2281 gaaaacgctg ggcgttaatc agagaggcga attatgtgtc agaggaccta tgattatgtc
     2341 cggttatgta aacaatccgg aagcgaccaa cgccttgatt gacaaggatg gatggctaca
     2401 ttctggagac atagcttact gggacgaaga cgaacacttc ttcatagttg accgcttgaa
     2461 gtctttaatt aaatacaaag gataccaggt ggcccccgct gaattggagt cgatattgtt
     2521 acaacacccc aacatcttcg acgcgggcgt ggcaggtctt cccgacgatg acgccggtga
     2581 acttcccgcc gccgttgttg ttttggagca cggaaagacg atgacggaaa aagagatcgt
     2641 ggattacgtc gccagtcaag taacaaccgc gaaaaagttg cgcggaggag ttgtgtttgt
     2701 ggacgaagta ccgaaaggtc ttaccggaaa actcgacgca agaaaaatca gagagatcct
     2761 cataaaggcc aagaagggcg gaaagtccaa attgctcgag gactacaaag accatgacgg
     2821 tgattataaa gatcatgaca tcgactacaa ggatgacgat gacaagtagt gagggcccga
     2881 tatctcgaca atcaacctct ggattacaaa atttgtgaaa gattgactgg tattcttaac
     2941 tatgttgctc cttttacgct atgtggatac gctgctttaa tgcctttgta tcatgctatt
     3001 gcttcccgta tggctttcat tttctcctcc ttgtataaat cctggttgct gtctctttat
     3061 gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt gcactgtgtt tgctgacgca
     3121 acccccactg gttggggcat tgccaccacc tgtcagctcc tttccgggac tttcgctttc
     3181 cccctcccta ttgccacggc ggaactcatc gccgcctgcc ttgcccgctg ctggacaggg
     3241 gctcggctgt tgggcactga caattccgtg gtgttgtcgg ggaagctgac gtcctttcca
     3301 tggctgctcg cctgtgttgc cacctggatt ctgcgcggga cgtccttctg ctacgtccct
     3361 tcggccctca atccagcgga ccttccttcc cgcggcctgc tgccggctct gcggcctctt
     3421 ccgcgtcttc gccttcgccc tcagacgagt cggatctccc tttgggccgc ctccccgcct
     3481 ggaaacgggg gaggctaact gaaacacgga aggagacaat accggaagga acccgcgcta
     3541 tgacggcaat aaaaagacag aataaaacgc acgggtgttg ggtcgtttgt tcataaacgc
     3601 ggggttcggt cccagggctg gcactctgtc gataccccac cgagacccca ttggggccaa
     3661 tacgcccgcg tttcttcctt ttccccaccc caccccccaa gttcgggtga aggcccaggg
     3721 ctcgcagcca acgtcggggc ggcaggccct gccatagcag atctgcgcag ctggggctct
     3781 agggggtatc cccacgcgcc ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg
     3841 cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct
     3901 tcctttctcg ccacgttcgc cggctttccc cgtcaagctc taaatcgggg catcccttta
     3961 gggttccgat ttagtgcttt acggcacctc gaccccaaaa aacttgatta gggtgatggt
     4021 tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg
     4081 ttctttaata gtggactctt gttccaaact ggaacaacac tcaaccctat ctcggtctat
     4141 tcttttgatt tataagggat tttggggatt tcggcctatt ggttaaaaaa tgagctgatt
     4201 taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt
     4261 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     4321 ggtgtggaaa gtccccaggc tccccagcag gcagaagtat gcaaagcatg catctcaatt
     4381 agtcagcaac catagtcccg cccctaactc cgcccatccc gcccctaact ccgcccagtt
     4441 ccgcccattc tccgccccat ggctgactaa ttttttttat ttatgcagag gccgaggccg
     4501 cctctgcctc tgagctattc cagaagtagt gaggaggctt ttttggaggc ctaggctttt
     4561 gcaaaaagct cccgggagct tgtatatcca ttttcggatc tgatcaagag acaggatgag
     4621 gatcgtttcg catgattgaa caagatggat tgcacgcagg ttctccggcc gcttgggtgg
     4681 agaggctatt cggctatgac tgggcacaac agacaatcgg ctgctctgat gccgccgtgt
     4741 tccggctgtc agcgcagggg cgcccggttc tttttgtcaa gaccgacctg tccggtgccc
     4801 tgaatgaact gcaggacgag gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt
     4861 gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga ctggctgcta ttgggcgaag
     4921 tgccggggca ggatctcctg tcatctcacc ttgctcctgc cgagaaagta tccatcatgg
     4981 ctgatgcaat gcggcggctg catacgcttg atccggctac ctgcccattc gaccaccaag
     5041 cgaaacatcg catcgagcga gcacgtactc ggatggaagc cggtcttgtc gatcaggatg
     5101 atctggacga agagcatcag gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc
     5161 gcatgcccga cggcgaggat ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca
     5221 tggtggaaaa tggccgcttt tctggattca tcgactgtgg ccggctgggt gtggcggacc
     5281 gctatcagga catagcgttg gctacccgtg atattgctga agagcttggc ggcgaatggg
     5341 ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga ttcgcagcgc atcgccttct
     5401 atcgccttct tgacgagttc ttctgagcgg gactctgggg ttcgcgaaat gaccgaccaa
     5461 gcgacgccca acctgccatc acgagatttc gattccaccg ccgccttcta tgaaaggttg
     5521 ggcttcggaa tcgttttccg ggacgccggc tggatgatcc tccagcgcgg ggatctcatg
     5581 ctggagttct tcgcccaccc caacttgttt attgcagctt ataatggtta caaataaagc
     5641 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5701 tccaaactca tcaatgtatc ttatcatgtc tgtataccgt cgacctctag ctagagcttg
     5761 gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt atccgctcac aattccacac
     5821 aacatacgag ccggaagcat aaagtgtaaa gcctggggtg cctaatgagt gagctaactc
     5881 acattaattg cgttgcgctc actgcccgct ttccagtcgg gaaacctgtc gtgccagctg
     5941 cattaatgaa tcggccaacg cgcggggaga ggcggtttgc gtattgggcg ctcttccgct
     6001 tcctcgctca ctgactcgct gcgctcggtc gttcggctgc ggcgagcggt atcagctcac
     6061 tcaaaggcgg taatacggtt atccacagaa tcaggggata acgcaggaaa gaacatgtga
     6121 gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg cgttgctggc gtttttccat
     6181 aggctccgcc cccctgacga gcatcacaaa aatcgacgct caagtcagag gtggcgaaac
     6241 ccgacaggac tataaagata ccaggcgttt ccccctggaa gctccctcgt gcgctctcct
     6301 gttccgaccc tgccgcttac cggatacctg tccgcctttc tcccttcggg aagcgtggcg
     6361 ctttctcaat gctcacgctg taggtatctc agttcggtgt aggtcgttcg ctccaagctg
     6421 ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg ccttatccgg taactatcgt
     6481 cttgagtcca acccggtaag acacgactta tcgccactgg cagcagccac tggtaacagg
     6541 attagcagag cgaggtatgt aggcggtgct acagagttct tgaagtggtg gcctaactac
     6601 ggctacacta gaaggacagt atttggtatc tgcgctctgc tgaagccagt taccttcgga
     6661 aaaagagttg gtagctcttg atccggcaaa caaaccaccg ctggtagcgg tggttttttt
     6721 gtttgcaagc agcagattac gcgcagaaaa aaaggatctc aagaagatcc tttgatcttt
     6781 tctacggggt ctgacgctca gtggaacgaa aactcacgtt aagggatttt ggtcatgaga
     6841 ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa aatgaagttt taaatcaatc
     6901 taaagtatat atgagtaaac ttggtctgac agttaccaat gcttaatcag tgaggcacct
     6961 atctcagcga tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata
     7021 actacgatac gggagggctt accatctggc cccagtgctg caatgatacc gcgagaccca
     7081 cgctcaccgg ctccagattt atcagcaata aaccagccag ccggaagggc cgagcgcaga
     7141 agtggtcctg caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga
     7201 gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg
     7261 gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga
     7321 gttacatgat cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt
     7381 gtcagaagta agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct
     7441 cttactgtca tgccatccgt aagatgcttt tctgtgactg gtgagtactc aaccaagtca
     7501 ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat
     7561 accgcgccac atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcga
     7621 aaactctcaa ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc
     7681 aactgatctt cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg
     7741 caaaatgccg caaaaaaggg aataagggcg acacggaaat gttgaatact catactcttc
     7801 ctttttcaat attattgaag catttatcag ggttattgtc tcatgagcgg atacatattt
     7861 gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg aaaagtgcca
     7921 cctgacgtcg acggatcggg agatctcccg atcccctatg gtcgactctc agtacaatct
     7981 gctctgatgc cgcatagtta agccagtatc tgctccctgc ttgtgtgttg gaggtcgctg
     8041 agtagtgcgc gagcaaaatt taagctacaa caaggcaagg cttgaccgac aattgcatga
     8101 agaatctgct tagg
//
