LOCUS       V022232                 8807 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022232
VERSION     V022232
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             683..3454
                     /label="ATG9B(NM_001317056)"
                     /note="ATG9B(NM_001317056)"
                     /gene="ATG9B"
     CDS             3494..3559
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    3583..4171
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    4237..4285
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      4487..4915
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        4929..5258
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             5325..6119
                     /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    6295..6416
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(6465..6481)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    6489..6505
                     /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(6513..6543)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    6558..6579
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(6867..7455)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(7626..8486)
                     /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(8487..8591)
                     /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 ggatccgcca ccatggtgag ccgaatgggc tgggggggga gaagaaggcg
      721 gctggggcgg tggggagatc tggggcccgg atcggtgccc ctcctcccca tgccactgcc
      781 acctcctcct cctccttcat gccggggacc tgggggaggg aggatctcca tcttctctct
      841 gtcccctgcc cctcatacaa gaagctcccc ctcctcattt tcccctccca ccgcagggcc
      901 cccttgctca gtgctacagg ggacaggggc ttctcagtct tgccacagtg ctctccctat
      961 cccagccacc cccccaacac aggctcaacc tgcaatgaca cctgcctctg catctccctc
     1021 ctggggatcc cactccaccc cacccctggc cccggcaacc cccactccct cacagcagtg
     1081 cccccaggac tctcctgggc tgcgggtagg ccctttgatc cctgaacagg attatgagcg
     1141 gctggaggac tgtgaccctg aggggtccca agactcaccc atccacgggg aggagcagca
     1201 acccctgctt catgtccctg aagggctccg aggctcctgg catcacatcc agaacctgga
     1261 cagtttcttc accaagatct acagctacca ccagcggaat ggctttgcct gcatcttgct
     1321 ggaggatgtc ttccagctgg gacaatttat tttcattgtc accttcacaa ccttcctcct
     1381 tcgatgcgtg gattacaatg ttctctttgc caaccaacca agtaaccata ccagacctgg
     1441 gccgttccac agcaaagtga ccctgtcaga tgccatccta ccctcagccc agtgtgctga
     1501 gaggatccgc tccagcccgc tgctggtcct cctcctggtc ctggctgccg gcttctggct
     1561 ggtccaactg cttcgctcag tctgcaacct cttcagctac tgggacatcc aggtgtttta
     1621 cagggaggcc ctgcacatcc ccccggagga gctgagctcg gttccctggg cagaggtgca
     1681 gtcccgcctc ttggcactgc agcggagcgg gggcctgtgc gtgcagccgc ggcccctgac
     1741 ggagctggac atccaccacc gcatcctgcg ctacaccaac taccaggtgg cgctggccaa
     1801 caaaggcctg ctgccggccc gctgcccgct gccctgggga ggcagtgcgg ctttcctcag
     1861 ccgcggcctg gcgctcaatg tcgacctgct gctcttccgc ggtcccttct cgctcttccg
     1921 cgggggctgg gagctgccgc acgcctacaa gcgcagcgac cagcggggcg ccctagcagc
     1981 gcgctggggg cgcacagtgc tgctgctggc cgccctgaac ctggcgctga gcccgctggt
     2041 gctggcctgg caggttctgc acgtcttcta tagccacgtg gagctgctgc ggcgcgagcc
     2101 tggcgcgctg ggggcgcgcg gctggtcccg cctggcgcgc ttgcagctgc gccacttcaa
     2161 cgagctgccg cacgagctgc gcgcgcgcct ggcccgcgcc taccgccccg ccgccgcctt
     2221 cctgcgcacc gctgcgcccc ccgcgcccct gcgcacgctg ctggcccgcc agctcgtttt
     2281 cttcgcgggt gcactcttcg ccgcgctgct tgtgctcacc gtctacgacg aggacgtgct
     2341 agccgtggag cacgtgctca ccgccatgac cgcgctcggg gtcaccgcca ccgtcgccag
     2401 gtctttcatt ccggaagagc agtgccaggg tcgtgcgccg cagctcctgc tgcagacagc
     2461 cctggcccac atgcactacc tcccggagga gcccggcccc ggcggcaggg accgcgccta
     2521 ccggcagatg gcgcagctgc tgcagtaccg agcggtctcc ctcctggagg agctcctgtc
     2581 cccgctcctc accccgctgt ttctgctttt ctggttccgc cctcgtgccc tggagattat
     2641 cgactttttt catcacttca ctgtggatgt ggctggggtt ggggacatct gttcctttgc
     2701 ccttatggat gtgaagcgcc acggacaccc tcagtggctc tcggcgggac agactgaggc
     2761 ctcgctgtct cagcgtgcgg aggacggcaa gactgagctt tctttgatgc ggttctccct
     2821 ggcgcatcca ctctggcgcc ccccagggca cagctctaag tttcttgggc acctctgggg
     2881 ccgagtacaa caagatgcag ctgcctgggg tgccacctcg gctcgcggcc cctccacccc
     2941 gggggtgctc agcaactgca cctcgcccct gcctgaggcc ttcctggcca acctcttcgt
     3001 gcaccctctc ctgcctccga gagatctgag cccgacagcc ccctgtccag ctgcggccac
     3061 agccagcctc cttgcctcca tttcccgaat tgcccaggac cccagctctg tgtccccagg
     3121 aggcactggg ggccagaagc tggcccagct cccagaactt gcttctgccg agatgagtct
     3181 ccatgtcatc tacctgcacc agcttcacca gcagcagcag cagcaggagc cgtggggtga
     3241 ggctgcagcc tccatcctgt ccaggccctg ctccagcccc tcacagccac cctcgcctga
     3301 tgaggagaag ccatcctggt caagtgacgg ctccagtcct gcctctagcc ccagacaaca
     3361 gtggggaacc cagaaggccc ggaatctgtt ccccggaggg tttcaggtga ccacagacac
     3421 ccagaaggag cctgaccggg cctcttgcac tgacgaattc tgcagatatc cagcacagtg
     3481 gcggccgctc gaggactaca aagaccatga cggtgattat aaagatcatg acatcgacta
     3541 caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc tctggattac
     3601 aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga
     3661 tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt cattttctcc
     3721 tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt tgtcaggcaa
     3781 cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg cattgccacc
     3841 acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac ggcggaactc
     3901 atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac tgacaattcc
     3961 gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt tgccacctgg
     4021 attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc ggaccttcct
     4081 tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg ccctcagacg
     4141 agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta actgaaacac
     4201 ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga cagaataaaa
     4261 cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg ctggcactct
     4321 gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc cttttcccca
     4381 ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg ggcggcaggc
     4441 cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc gccctgtagc
     4501 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     4561 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     4621 ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc tttacggcac
     4681 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     4741 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4801 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgggg
     4861 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4921 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4981 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     5041 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     5101 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     5161 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     5221 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat
     5281 ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt gaacaagatg
     5341 gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat gactgggcac
     5401 aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag gggcgcccgg
     5461 ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac gaggcagcgc
     5521 ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac gttgtcactg
     5581 aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc ctgtcatctc
     5641 accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg ctgcatacgc
     5701 ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag cgagcacgta
     5761 ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat caggggctcg
     5821 cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag gatctcgtcg
     5881 tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc ttttctggat
     5941 tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg ttggctaccc
     6001 gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg ctttacggta
     6061 tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag ttcttctgag
     6121 cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc atcacgagat
     6181 ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt ccgggacgcc
     6241 ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca ccccaacttg
     6301 tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa
     6361 gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat
     6421 gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata gctgtttcct
     6481 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     6541 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     6601 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     6661 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     6721 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     6781 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     6841 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     6901 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     6961 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     7021 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg ctgtaggtat
     7081 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     7141 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     7201 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     7261 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt
     7321 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     7381 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     7441 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     7501 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     7561 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     7621 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     7681 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     7741 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     7801 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     7861 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     7921 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     7981 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     8041 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     8101 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     8161 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     8221 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     8281 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     8341 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     8401 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     8461 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     8521 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     8581 ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc gggagatctc
     8641 ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag ttaagccagt
     8701 atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa atttaagcta
     8761 caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
