LOCUS       V030338                 8134 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V030338
VERSION     V030338
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        235..614
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        615..818
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        863..881
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             937..4086
                     /label="ULK1(NM_003565)"
                     /note="ULK1(NM_003565)"
                     /gene="ULK1"
     CDS             4099..4164
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     polyA_signal    4199..4423
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4469..4897
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4911..5240
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5252..5650
                     /codon_start=1
                     /gene="Aspergillus terreus BSD"
                     /note="confers resistance to blasticidin"
                     /product="blasticidin S deaminase"
                     /transl_table=1
                     /translation="MAKPLSQEESTLIERATATINSIPISEDYSVASAALSSDGRIFTG
                     VNVYHFTGGPCAELVVLGTAAAAAAGNLTCIVAIGNENRGILSPCGRCRQVLLDLHPGI
                     KAIVKDSDGQPTAVGIRELLPSGYVWEG*"
                     /label="BSD"
     polyA_signal    5810..5931
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5980..5996)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6004..6020
                     /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(6028..6058)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6073..6094
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6382..6967)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7138..7998)
                     /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(7999..8103)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gacggatcgg gagatctccc gatcccctat ggtgcactct cagtacaatc tgctctgatg
       61 ccgcatagtt aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg
      121 cgagcaaaat ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc
      181 ttagggttag gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt
      241 gattattgac tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata
      301 tggagttccg cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc
      361 cccgcccatt gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc
      421 attgacgtca atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt
      481 atcatatgcc aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt
      541 atgcccagta catgacctta tgggactttc ctacttggca gtacatctac gtattagtca
      601 tcgctattac catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg
      661 actcacgggg atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc
      721 aaaatcaacg ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg
      781 gtaggcgtgt acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca
      841 ctgcttactg gcttatcgaa attaatacga ctcactatag ggagacccaa gctggctagc
      901 aagctttcta gagaattcga tatcggatcc gccaccatgg agcccggccg cggcggcaca
      961 gagaccgtgg gcaagttcga gttctcccgc aaggacctga tcggccacgg cgccttcgcg
     1021 gtggtcttca agggccgcca ccgcgagaag cacgatttgg aggtcgccgt caagtgcatt
     1081 aacaagaaga acctcgccaa gtctcagacg ctgctgggga aggaaatcaa aatcctgaag
     1141 gaactgaaac atgaaaacat cgtggccctg tacgacttcc aggaaatggc taattctgtc
     1201 tacctggtta tggagtactg caacggtggg gacctggccg actacctgca cgccatgcgc
     1261 acgctgagcg aggacaccat caggctcttc ctgcagcaga tcgcgggcgc catgcggctt
     1321 ctgcacagca aaggcatcat ccaccgcgac ctgaaaccgc agaacatcct gctgtccaac
     1381 cccgccggcc gccgcgccaa ccccaacagc atccgcgtca agatcgctga cttcggcttc
     1441 gcgcggtacc tccagagcaa catgatggcg gccacactct gcggctcccc catgtacatg
     1501 gcccccgagg tcatcatgtc ccagcactac gacgggaagg cggacctgtg gagcatcggc
     1561 accatcgtct accagtgcct gacggggaag gcgcccttcc aggccagcag cccccaggac
     1621 ctgcgcctgt tctacgagaa gaacaagacg ttggtcccca ccatcccccg ggagacctcg
     1681 gccccgctgc ggcagctgct cctggcccta ctgcaacgca accacaagga ccgcatggac
     1741 ttcgatgagt tttttcatca ccctttcctc gatgccagcc cctcggtcag gaaatcccca
     1801 cccgtgcctg tgccctcgta cccaagctcg gggtccggca gcagctccag cagcagctcc
     1861 acctcccacc tggcctcccc gccgtccctg ggcgagatgc agcagctgca gaagaccctg
     1921 gcctccccgg ctgacaccgc tggcttcctg cacagctccc gggactctgg tggcagcaag
     1981 gactcttcct gtgacacaga cgacttcgtc atggtccccg cgcagtttcc aggtgacctg
     2041 gtggctgagg cgcccagtgc caaacccccg ccagacagcc tgatgtgcag tgggagctca
     2101 ctggtggcct ctgcgggctt ggagagccac ggccggaccc catctccatc cccaccctgc
     2161 agcagctccc ccagtccctc aggccgggct ggcccgttct ccagcagcag gtgcggcgcc
     2221 tctgtcccca tcccagtccc cacgcaggtg cagaactacc agcgcattga gcgaaacctg
     2281 cagtcaccca cccagttcca aacacctcgg tcctctgcca tccgcaggtc aggcagcacc
     2341 agccccctgg gctttgcaag ggccagcccc tcgccccctg cccacgctga gcatggaggc
     2401 gtcctggcca ggaagatgtc tctgggtgga ggccggccct acacgccatc tcctcaagtt
     2461 ggaaccatcc ctgagcggcc aggctggagc gggacgccct ccccacaggg agctgagatg
     2521 cggggtggca ggtcccctcg tccaggctcc tctgcacccg agcactctcc ccgcacttcc
     2581 gggctgggct gccgcctgca cagcgccccc aacctgtctg acttgcacgt cgtccgcccc
     2641 aagctgccca aaccccccac ggaccccctg ggagctgtgt tcagcccacc acaggccagc
     2701 cctccccagc cgtcccacgg cctgcagtcc tgccggaacc tgcggggctc acccaagctg
     2761 cccgacttcc tgcagcgaaa ccccctgccc cccatcctgg gctcccccac caaggctgtg
     2821 ccctcctttg acttcccgaa gacccccagc tcccagaacc tgctggccct cctagcccgg
     2881 cagggcgtgg tgatgacgcc ccctcgaaac cggacgctgc ccgacctctc ggaggtggga
     2941 cccttccatg gtcagccgtt gggccctggc ctgcggccag gcgaggaccc caagggcccc
     3001 tttggccggt ctttcagcac cagccgcctc actgacctgc tccttaaggc ggcgtttggg
     3061 acacaagccc cggacccggg cagcacggag agcctgcagg agaagcccat ggagatcgca
     3121 ccctcagctg gctttggagg gagcctgcac ccaggagccc gtgctggggg caccagcagc
     3181 ccttccccgg tggtcttcac cgtgggctct cccccgagcg ggagcacgcc cccccagggc
     3241 ccccgcacca ggatgttctc agcgggcccc actggctctg ccagctcttc tgcccgccac
     3301 ctggtgcctg ggccctgcag cgaggcccca gcccctgagc tccctgctcc aggacacggc
     3361 tgcagctttg ccgaccccat tactgcgaac ctggaggggg ctgtgacctt cgaggccccc
     3421 gacctccctg aggagaccct catggagcaa gagcacacgg agatcctgcg tggcctgcgc
     3481 ttcacgctgc tgttcgtgca gcacgtcctg gagatcgcag ccctgaaggg cagcgccagt
     3541 gaggcggcgg ggggccctga gtaccagctg caggagagtg tggtggccga ccagatcagc
     3601 ctgctgagcc gagaatgggg cttcgcggaa cagctggtgc tgtacctgaa ggtggccgag
     3661 ctactgtcct ccggcctgca aagtgccatc gaccagatcc gggccggcaa gctctgcctg
     3721 tcgtccactg tgaagcaggt ggtgcgcagg ctgaatgagc tgtacaaggc cagcgtggtg
     3781 tcctgccagg gcctgagcct gcggctgcag cgcttcttcc tggacaagca gcggctcctg
     3841 gaccgcattc acagcatcac tgccgagagg ctcatcttca gccacgctgt gcagatggtg
     3901 cagtcggctg ccctggacga gatgttccag caccgtgagg gctgcgtccc acgctaccac
     3961 aaggccctgc tgctcctgga ggggctgcag cacatgctct cggaccaggc cgacatcgag
     4021 aacgtcacca agtgcaagct gtgcattgag cggagactct cggcgctgct gactggcatc
     4081 tgtgccctcg agaccggtga ctacaaagac catgacggtg attataaaga tcatgacatc
     4141 gattacaagg atgacgatga caagtgaccg gtaagtttaa accgctgatc agcctcgact
     4201 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     4261 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     4321 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     4381 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     4441 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     4501 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     4561 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     4621 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     4681 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     4741 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     4801 cctatctcgg tctattcttt tgatttataa gggattttgg ccatttcggc ctattggtta
     4861 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     4921 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     4981 attagtcagc aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     5041 gcatgcatct caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc
     5101 taactccgcc cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg
     5161 cagaggccga ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg
     5221 gaggcctagg cttttgcaaa aagctcccgg gatggccaag cctttgtctc aagaagaatc
     5281 caccctcatt gaaagagcaa cggctacaat caacagcatc cccatctctg aagactacag
     5341 cgtcgccagc gcagctctct ctagcgacgg ccgcatcttc actggtgtca atgtatatca
     5401 ttttactggg ggaccttgtg cagaactcgt ggtgctgggc actgctgctg ctgcggcagc
     5461 tggcaacctg acttgtatcg tcgcgatcgg aaatgagaac aggggcatct tgagcccctg
     5521 cggacggtgc cgacaggtgc ttctcgatct gcatcctggg atcaaagcca tagtgaagga
     5581 cagtgatgga cagccgacgg cagttgggat tcgtgaattg ctgccctctg gttatgtgtg
     5641 ggagggctaa ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac gagatttcga
     5701 ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg acgccggctg
     5761 gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca acttgtttat
     5821 tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5881 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5941 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     6001 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     6061 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     6121 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     6181 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6241 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6301 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6361 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6421 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6481 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6541 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6601 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6661 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6721 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6781 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6841 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6901 aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6961 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     7021 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     7081 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     7141 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     7201 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     7261 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     7321 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     7381 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     7441 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     7501 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7561 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7621 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7681 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7741 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7801 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7861 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7921 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7981 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     8041 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     8101 gcgcacattt ccccgaaaag tgccacctga cgtc
//
