LOCUS       V028042                 8597 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V028042
VERSION     V028042
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        202..581
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        582..785
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        830..848
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             908..4057
                     /label="ULK1(NM_003565)"
                     /note="ULK1(NM_003565)"
                     /gene="ULK1"
     CDS             4058..4084
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     polyA_signal    4164..4388
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      4434..4862
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4876..5205
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5272..6066
                     /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    6240..6361
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6410..6426)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6434..6450
                     /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(6458..6488)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6503..6524
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6812..7397)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7568..8428)
                     /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(8429..8533)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg ctccctgctt
       61 gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca aggcaaggct
      121 tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg cttcgcgatg
      181 tacgggccag atatacgcgt tgacattgat tattgactag ttattaatag taatcaatta
      241 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
      301 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
      361 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
      421 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
      481 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
      541 cttggcagta catctacgta ttagtcatcg ctattaccat ggtgatgcgg ttttggcagt
      601 acatcaatgg gcgtggatag cggtttgact cacggggatt tccaagtctc caccccattg
      661 acgtcaatgg gagtttgttt tggcaccaaa atcaacggga ctttccaaaa tgtcgtaaca
      721 actccgcccc attgacgcaa atgggcggta ggcgtgtacg gtgggaggtc tatataagca
      781 gagctctctg gctaactaga gaacccactg cttactggct tatcgaaatt aatacgactc
      841 actataggga gacccaagct ggctagcgtt taaacttaag cttggtaccg agctcggatc
      901 cgccaccatg gagcccggcc gcggcggcac agagaccgtg ggcaagttcg agttctcccg
      961 caaggacctg atcggccacg gcgccttcgc ggtggtcttc aagggccgcc accgcgagaa
     1021 gcacgatttg gaggtcgccg tcaagtgcat taacaagaag aacctcgcca agtctcagac
     1081 gctgctgggg aaggaaatca aaatcctgaa ggaactgaaa catgaaaaca tcgtggccct
     1141 gtacgacttc caggaaatgg ctaattctgt ctacctggtt atggagtact gcaacggtgg
     1201 ggacctggcc gactacctgc acgccatgcg cacgctgagc gaggacacca tcaggctctt
     1261 cctgcagcag atcgcgggcg ccatgcggct tctgcacagc aaaggcatca tccaccgcga
     1321 cctgaaaccg cagaacatcc tgctgtccaa ccccgccggc cgccgcgcca accccaacag
     1381 catccgcgtc aagatcgctg acttcggctt cgcgcggtac ctccagagca acatgatggc
     1441 ggccacactc tgcggctccc ccatgtacat ggcccccgag gtcatcatgt cccagcacta
     1501 cgacgggaag gcggacctgt ggagcatcgg caccatcgtc taccagtgcc tgacggggaa
     1561 ggcgcccttc caggccagca gcccccagga cctgcgcctg ttctacgaga agaacaagac
     1621 gttggtcccc accatccccc gggagacctc ggccccgctg cggcagctgc tcctggccct
     1681 actgcaacgc aaccacaagg accgcatgga cttcgatgag ttttttcatc accctttcct
     1741 cgatgccagc ccctcggtca ggaaatcccc acccgtgcct gtgccctcgt acccaagctc
     1801 ggggtccggc agcagctcca gcagcagctc cacctcccac ctggcctccc cgccgtccct
     1861 gggcgagatg cagcagctgc agaagaccct ggcctccccg gctgacaccg ctggcttcct
     1921 gcacagctcc cgggactctg gtggcagcaa ggactcttcc tgtgacacag acgacttcgt
     1981 catggtcccc gcgcagtttc caggtgacct ggtggctgag gcgcccagtg ccaaaccccc
     2041 gccagacagc ctgatgtgca gtgggagctc actggtggcc tctgcgggct tggagagcca
     2101 cggccggacc ccatctccat ccccaccctg cagcagctcc cccagtccct caggccgggc
     2161 tggcccgttc tccagcagca ggtgcggcgc ctctgtcccc atcccagtcc ccacgcaggt
     2221 gcagaactac cagcgcattg agcgaaacct gcagtcaccc acccagttcc aaacacctcg
     2281 gtcctctgcc atccgcaggt caggcagcac cagccccctg ggctttgcaa gggccagccc
     2341 ctcgccccct gcccacgctg agcatggagg cgtcctggcc aggaagatgt ctctgggtgg
     2401 aggccggccc tacacgccat ctcctcaagt tggaaccatc cctgagcggc caggctggag
     2461 cgggacgccc tccccacagg gagctgagat gcggggtggc aggtcccctc gtccaggctc
     2521 ctctgcaccc gagcactctc cccgcacttc cgggctgggc tgccgcctgc acagcgcccc
     2581 caacctgtct gacttgcacg tcgtccgccc caagctgccc aaacccccca cggaccccct
     2641 gggagctgtg ttcagcccac cacaggccag ccctccccag ccgtcccacg gcctgcagtc
     2701 ctgccggaac ctgcggggct cacccaagct gcccgacttc ctgcagcgaa accccctgcc
     2761 ccccatcctg ggctccccca ccaaggctgt gccctccttt gacttcccga agacccccag
     2821 ctcccagaac ctgctggccc tcctagcccg gcagggcgtg gtgatgacgc cccctcgaaa
     2881 ccggacgctg cccgacctct cggaggtggg acccttccat ggtcagccgt tgggccctgg
     2941 cctgcggcca ggcgaggacc ccaagggccc ctttggccgg tctttcagca ccagccgcct
     3001 cactgacctg ctccttaagg cggcgtttgg gacacaagcc ccggacccgg gcagcacgga
     3061 gagcctgcag gagaagccca tggagatcgc accctcagct ggctttggag ggagcctgca
     3121 cccaggagcc cgtgctgggg gcaccagcag cccttccccg gtggtcttca ccgtgggctc
     3181 tcccccgagc gggagcacgc ccccccaggg cccccgcacc aggatgttct cagcgggccc
     3241 cactggctct gccagctctt ctgcccgcca cctggtgcct gggccctgca gcgaggcccc
     3301 agcccctgag ctccctgctc caggacacgg ctgcagcttt gccgacccca ttgctgcgaa
     3361 cctggagggg gctgtgacct tcgaggcccc cgacctccct gaggagaccc tcatggagca
     3421 agagcacacg gagatcctgc gtggcctgcg cttcacgctg ctgttcgtgc agcacgtcct
     3481 ggagatcgca gccctgaagg gcagcgccag tgaggcggcg gggggccctg agtaccagct
     3541 gcaggagagt gtggtggccg accagatcag cctgctgagc cgagaatggg gcttcgcgga
     3601 acagctggtg ctgtacctga aggtggccga gctactgtcc tccggcctgc aaagtgccat
     3661 cgaccagatc cgggccggca agctctgcct gtcgtccact gtgaagcagg tggtgcgcag
     3721 gctgaatgag ctgtacaagg ccagcgtggt gtcctgccag ggcctgagcc tgcggctgca
     3781 gcgcttcttc ctggacaagc agcggctcct ggaccgcatt cacagcatca ctgccgagag
     3841 gctcatcttc agccacgctg tgcagatggt gcagtcggct gccctggacg agatgttcca
     3901 gcaccgtgag ggctgcgtcc cacgctacca caaggccctg ctgctcctgg aggggctgca
     3961 gcacatgctc tcggaccagg ccgacatcga gaacgtcacc aagtgcaagc tgtgcattga
     4021 gcggagactc tcggcgctgc tgactggcat ctgtgcctac ccatacgatg ttccagatta
     4081 cgcttgagaa ttctgcagat atccagcaca gtggcggccg ctcgagtcta gagggcccgt
     4141 ttaaacccgc tgatcagcct cgactgtgcc ttctagttgc cagccatctg ttgtttgccc
     4201 ctcccccgtg ccttccttga ccctggaagg tgccactccc actgtccttt cctaataaaa
     4261 tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct attctggggg gtggggtggg
     4321 gcaggacagc aagggggagg attgggaaga caatagcagg catgctgggg atgcggtggg
     4381 ctctatggct tctgaggcgg aaagaaccag ctggggctct agggggtatc cccacgcgcc
     4441 ctgtagcggc gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact
     4501 tgccagcgcc ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc
     4561 cggctttccc cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt
     4621 acggcacctc gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc
     4681 ctgatagacg gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt
     4741 gttccaaact ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat
     4801 tttgccgatt tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa
     4861 ttaattctgt ggaatgtgtg tcagttaggg tgtggaaagt ccccaggctc cccagcaggc
     4921 agaagtatgc aaagcatgca tctcaattag tcagcaacca ggtgtggaaa gtccccaggc
     4981 tccccagcag gcagaagtat gcaaagcatg catctcaatt agtcagcaac catagtcccg
     5041 cccctaactc cgcccatccc gcccctaact ccgcccagtt ccgcccattc tccgccccat
     5101 ggctgactaa ttttttttat ttatgcagag gccgaggccg cctctgcctc tgagctattc
     5161 cagaagtagt gaggaggctt ttttggaggc ctaggctttt gcaaaaagct cccgggagct
     5221 tgtatatcca ttttcggatc tgatcaagag acaggatgag gatcgtttcg catgattgaa
     5281 caagatggat tgcacgcagg ttctccggcc gcttgggtgg agaggctatt cggctatgac
     5341 tgggcacaac agacaatcgg ctgctctgat gccgccgtgt tccggctgtc agcgcagggg
     5401 cgcccggttc tttttgtcaa gaccgacctg tccggtgccc tgaatgaact gcaggacgag
     5461 gcagcgcggc tatcgtggct ggccacgacg ggcgttcctt gcgcagctgt gctcgacgtt
     5521 gtcactgaag cgggaaggga ctggctgcta ttgggcgaag tgccggggca ggatctcctg
     5581 tcatctcacc ttgctcctgc cgagaaagta tccatcatgg ctgatgcaat gcggcggctg
     5641 catacgcttg atccggctac ctgcccattc gaccaccaag cgaaacatcg catcgagcga
     5701 gcacgtactc ggatggaagc cggtcttgtc gatcaggatg atctggacga agagcatcag
     5761 gggctcgcgc cagccgaact gttcgccagg ctcaaggcgc gcatgcccga cggcgaggat
     5821 ctcgtcgtga cccatggcga tgcctgcttg ccgaatatca tggtggaaaa tggccgcttt
     5881 tctggattca tcgactgtgg ccggctgggt gtggcggacc gctatcagga catagcgttg
     5941 gctacccgtg atattgctga agagcttggc ggcgaatggg ctgaccgctt cctcgtgctt
     6001 tacggtatcg ccgctcccga ttcgcagcgc atcgccttct atcgccttct tgacgagttc
     6061 ttctgagcgg gactctgggg ttcgaaatga ccgaccaagc gacgcccaac ctgccatcac
     6121 gagatttcga ttccaccgcc gccttctatg aaaggttggg cttcggaatc gttttccggg
     6181 acgccggctg gatgatcctc cagcgcgggg atctcatgct ggagttcttc gcccacccca
     6241 acttgtttat tgcagcttat aatggttaca aataaagcaa tagcatcaca aatttcacaa
     6301 ataaagcatt tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt
     6361 atcatgtctg tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt
     6421 ttcctgtgtg aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa
     6481 agtgtaaagc ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac
     6541 tgcccgcttt ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg
     6601 cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc
     6661 gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat
     6721 ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca
     6781 ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc
     6841 atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc
     6901 aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg
     6961 gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta
     7021 ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg
     7081 ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac
     7141 acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag
     7201 gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat
     7261 ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat
     7321 ccggcaaaca aaccaccgct ggtagcggtt tttttgtttg caagcagcag attacgcgca
     7381 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     7441 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     7501 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     7561 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     7621 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     7681 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     7741 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     7801 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     7861 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     7921 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     7981 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     8041 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     8101 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     8161 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     8221 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     8281 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     8341 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     8401 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     8461 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     8521 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     8581 tcccgatccc ctatggt
//
