LOCUS       V044273                 7474 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V044273
VERSION     V044273
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             706..2130
                     /label="Tfeb(NM_001161723)"
                     /note="Tfeb(NM_001161723)"
                     /gene="Tfeb"
     CDS             2137..2226
                     /codon_start=1
                     /product="3 tandem Myc epitope tags"
                     /transl_table=1
                     /translation="EQKLISEEDLEQKLISEEDLEQKLISEEDL"
                     /label="3xMyc"
     misc_feature    2250..2838
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2904..2952
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3154..3582
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3596..3925
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3992..4786
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    4962..5083
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5132..5148)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5156..5172
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(5180..5210)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5225..5246
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5534..6122)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6293..7153)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
     promoter        complement(7154..7258)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctggcta gcgtttaaac ttaagcttgg
      661 taccgagctc ggatccacta gtccagtgtg gtggaattcg ccaccatggc gtcacgcatc
      721 gggctgcgca tgcagctcat gcgggagcag gcccagcagg aggagcagcg agagcgcatg
      781 cagcagcagg ctgtcatgca ttatatgcaa cagcagcagc agcagcagca gcagctgggt
      841 gggcccccca ccccagccat caacacccct gtccacttcc agtcgccccc gcctgtgccc
      901 ggggaggtgc tgaaggtgca gtcctacctg gagaacccca cctcctacca cctgcaacag
      961 tcccagcatc agaaggttcg ggagtatctg tctgagacct atgggaacaa gtttgctgcc
     1021 cacgtgagcc cagcccaagg ttccccgaag cctgccccag cagcatcccc aggggtgcgg
     1081 gctggacacg tactgtccac ctcggccggc aacagtgctc ccaacagtcc catggccatg
     1141 ctacatatca gctccaaccc cgagaaagag tttgatgatg tcattgacaa cattatgcgc
     1201 ctggacagcg tgctgggcta catcaaccct gagatgcaga tgcctaacac gctgcccctg
     1261 tctagcagcc acctgaacgt gtacagcggt gacccccagg tcacagcctc catggtgggt
     1321 gtcaccagca gctcctgccc tgccgacctg actcagaagc gagagctaac agatgctgag
     1381 agcagagccc tggccaagga gcggcagaag aaagacaatc acaacctaat tgagagaaga
     1441 cgcaggttca acatcaatga ccggatcaag gagctgggaa tgctgatccc caaggccaac
     1501 gacctggacg tgcgctggaa caaaggcacc atcctcaagg cctctgtgga ttacatccgg
     1561 aggatgcaga aggacctgca gaagtcccgg gagctggaga accactcccg gcgcctggag
     1621 atgactaaca agcagctctg gctccgcatc caggagctgg agatgcaggc acgcgtgcac
     1681 ggcctcccca ccacctcgcc gtcgggtgtg aatatggccg agctggccca gcaggtggtg
     1741 aagcaagagt tgcccagtga ggatggccca ggggaggcgc tgatgctggg gcctgaggtc
     1801 cctgagcctg agcaaatgcc ggctcttcct ccccaggctc cgctgccctc ggccgcccag
     1861 ccacagtctc cgttccatca cctggacttc agccatggcc tgagctttgg gggtgggggc
     1921 gacgaggggc ccacaggtta ccccgatacc ctggggacag agcacggctc cccattcccc
     1981 aacctgtcca agaaggatct ggacttaatg ctcctagatg actccctgct ccccctggcc
     2041 tctgaccccc tcttttctac catgtctcct gaggcctcca aggccagcag ccgccggagc
     2101 agcttcagca tggaggaggg tgatgttctg ctcgaggaac aaaaactaat atcagaggaa
     2161 gatttggaac agaaactaat cagtgaagaa gatttagagc aaaagttaat ttccgaggag
     2221 gacttatagt gagggcccga tatctcgaca atcaacctct ggattacaaa atttgtgaaa
     2281 gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac gctgctttaa
     2341 tgcctttgta tcatgctatt gcttcccgta tggctttcat tttctcctcc ttgtataaat
     2401 cctggttgct gtctctttat gaggagttgt ggcccgttgt caggcaacgt ggcgtggtgt
     2461 gcactgtgtt tgctgacgca acccccactg gttggggcat tgccaccacc tgtcagctcc
     2521 tttccgggac tttcgctttc cccctcccta ttgccacggc ggaactcatc gccgcctgcc
     2581 ttgcccgctg ctggacaggg gctcggctgt tgggcactga caattccgtg gtgttgtcgg
     2641 ggaagctgac gtcctttcca tggctgctcg cctgtgttgc cacctggatt ctgcgcggga
     2701 cgtccttctg ctacgtccct tcggccctca atccagcgga ccttccttcc cgcggcctgc
     2761 tgccggctct gcggcctctt ccgcgtcttc gccttcgccc tcagacgagt cggatctccc
     2821 tttgggccgc ctccccgcct ggaaacgggg gaggctaact gaaacacgga aggagacaat
     2881 accggaagga acccgcgcta tgacggcaat aaaaagacag aataaaacgc acgggtgttg
     2941 ggtcgtttgt tcataaacgc ggggttcggt cccagggctg gcactctgtc gataccccac
     3001 cgagacccca ttggggccaa tacgcccgcg tttcttcctt ttccccaccc caccccccaa
     3061 gttcgggtga aggcccaggg ctcgcagcca acgtcggggc ggcaggccct gccatagcag
     3121 atctgcgcag ctggggctct agggggtatc cccacgcgcc ctgtagcggc gcattaagcg
     3181 cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc ctagcgcccg
     3241 ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc cgtcaagctc
     3301 taaatcgggg catcccttta gggttccgat ttagtgcttt acggcacctc gaccccaaaa
     3361 aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg gtttttcgcc
     3421 ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact ggaacaacac
     3481 tcaaccctat ctcggtctat tcttttgatt tataagggat tttggggatt tcggcctatt
     3541 ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttaattctgt ggaatgtgtg
     3601 tcagttaggg tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca
     3661 tctcaattag tcagcaacca ggtgtggaaa gtccccaggc tccccagcag gcagaagtat
     3721 gcaaagcatg catctcaatt agtcagcaac catagtcccg cccctaactc cgcccatccc
     3781 gcccctaact ccgcccagtt ccgcccattc tccgccccat ggctgactaa ttttttttat
     3841 ttatgcagag gccgaggccg cctctgcctc tgagctattc cagaagtagt gaggaggctt
     3901 ttttggaggc ctaggctttt gcaaaaagct cccgggagct tgtatatcca ttttcggatc
     3961 tgatcaagag acaggatgag gatcgtttcg catgattgaa caagatggat tgcacgcagg
     4021 ttctccggcc gcttgggtgg agaggctatt cggctatgac tgggcacaac agacaatcgg
     4081 ctgctctgat gccgccgtgt tccggctgtc agcgcagggg cgcccggttc tttttgtcaa
     4141 gaccgacctg tccggtgccc tgaatgaact gcaggacgag gcagcgcggc tatcgtggct
     4201 ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt gtcactgaag cgggaaggga
     4261 ctggctgcta ttgggcgaag tgccggggca ggatctcctg tcatctcacc ttgctcctgc
     4321 cgagaaagta tccatcatgg ctgatgcaat gcggcggctg catacgcttg atccggctac
     4381 ctgcccattc gaccaccaag cgaaacatcg catcgagcga gcacgtactc ggatggaagc
     4441 cggtcttgtc gatcaggatg atctggacga agagcatcag gggctcgcgc cagccgaact
     4501 gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat ctcgtcgtga cccatggcga
     4561 tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt tctggattca tcgactgtgg
     4621 ccggctgggt gtggcggacc gctatcagga catagcgttg gctacccgtg atattgctga
     4681 agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt tacggtatcg ccgctcccga
     4741 ttcgcagcgc atcgccttct atcgccttct tgacgagttc ttctgagcgg gactctgggg
     4801 ttcgcgaaat gaccgaccaa gcgacgccca acctgccatc acgagatttc gattccaccg
     4861 ccgccttcta tgaaaggttg ggcttcggaa tcgttttccg ggacgccggc tggatgatcc
     4921 tccagcgcgg ggatctcatg ctggagttct tcgcccaccc caacttgttt attgcagctt
     4981 ataatggtta caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac
     5041 tgcattctag ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tgtataccgt
     5101 cgacctctag ctagagcttg gcgtaatcat ggtcatagct gtttcctgtg tgaaattgtt
     5161 atccgctcac aattccacac aacatacgag ccggaagcat aaagtgtaaa gcctggggtg
     5221 cctaatgagt gagctaactc acattaattg cgttgcgctc actgcccgct ttccagtcgg
     5281 gaaacctgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga ggcggtttgc
     5341 gtattgggcg ctcttccgct tcctcgctca ctgactcgct gcgctcggtc gttcggctgc
     5401 ggcgagcggt atcagctcac tcaaaggcgg taatacggtt atccacagaa tcaggggata
     5461 acgcaggaaa gaacatgtga gcaaaaggcc agcaaaaggc caggaaccgt aaaaaggccg
     5521 cgttgctggc gtttttccat aggctccgcc cccctgacga gcatcacaaa aatcgacgct
     5581 caagtcagag gtggcgaaac ccgacaggac tataaagata ccaggcgttt ccccctggaa
     5641 gctccctcgt gcgctctcct gttccgaccc tgccgcttac cggatacctg tccgcctttc
     5701 tcccttcggg aagcgtggcg ctttctcaat gctcacgctg taggtatctc agttcggtgt
     5761 aggtcgttcg ctccaagctg ggctgtgtgc acgaaccccc cgttcagccc gaccgctgcg
     5821 ccttatccgg taactatcgt cttgagtcca acccggtaag acacgactta tcgccactgg
     5881 cagcagccac tggtaacagg attagcagag cgaggtatgt aggcggtgct acagagttct
     5941 tgaagtggtg gcctaactac ggctacacta gaaggacagt atttggtatc tgcgctctgc
     6001 tgaagccagt taccttcgga aaaagagttg gtagctcttg atccggcaaa caaaccaccg
     6061 ctggtagcgg tggttttttt gtttgcaagc agcagattac gcgcagaaaa aaaggatctc
     6121 aagaagatcc tttgatcttt tctacggggt ctgacgctca gtggaacgaa aactcacgtt
     6181 aagggatttt ggtcatgaga ttatcaaaaa ggatcttcac ctagatcctt ttaaattaaa
     6241 aatgaagttt taaatcaatc taaagtatat atgagtaaac ttggtctgac agttaccaat
     6301 gcttaatcag tgaggcacct atctcagcga tctgtctatt tcgttcatcc atagttgcct
     6361 gactccccgt cgtgtagata actacgatac gggagggctt accatctggc cccagtgctg
     6421 caatgatacc gcgagaccca cgctcaccgg ctccagattt atcagcaata aaccagccag
     6481 ccggaagggc cgagcgcaga agtggtcctg caactttatc cgcctccatc cagtctatta
     6541 attgttgccg ggaagctaga gtaagtagtt cgccagttaa tagtttgcgc aacgttgttg
     6601 ccattgctac aggcatcgtg gtgtcacgct cgtcgtttgg tatggcttca ttcagctccg
     6661 gttcccaacg atcaaggcga gttacatgat cccccatgtt gtgcaaaaaa gcggttagct
     6721 ccttcggtcc tccgatcgtt gtcagaagta agttggccgc agtgttatca ctcatggtta
     6781 tggcagcact gcataattct cttactgtca tgccatccgt aagatgcttt tctgtgactg
     6841 gtgagtactc aaccaagtca ttctgagaat agtgtatgcg gcgaccgagt tgctcttgcc
     6901 cggcgtcaat acgggataat accgcgccac atagcagaac tttaaaagtg ctcatcattg
     6961 gaaaacgttc ttcggggcga aaactctcaa ggatcttacc gctgttgaga tccagttcga
     7021 tgtaacccac tcgtgcaccc aactgatctt cagcatcttt tactttcacc agcgtttctg
     7081 ggtgagcaaa aacaggaagg caaaatgccg caaaaaaggg aataagggcg acacggaaat
     7141 gttgaatact catactcttc ctttttcaat attattgaag catttatcag ggttattgtc
     7201 tcatgagcgg atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca
     7261 catttccccg aaaagtgcca cctgacgtcg acggatcggg agatctcccg atcccctatg
     7321 gtcgactctc agtacaatct gctctgatgc cgcatagtta agccagtatc tgctccctgc
     7381 ttgtgtgttg gaggtcgctg agtagtgcgc gagcaaaatt taagctacaa caaggcaagg
     7441 cttgaccgac aattgcatga agaatctgct tagg
//
