LOCUS       V038669                 7723 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V038669
VERSION     V038669
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..2382
                     /label="Prkaa1(NM_001013367)"
                     /note="Prkaa1(NM_001013367)"
                     /gene="Prkaa1"
     CDS             2389..2415
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2419..2445
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     CDS             2449..2475
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /transl_table=1
                     /translation="YPYDVPDYA"
                     /label="HA"
     misc_feature    2499..3087
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3153..3201
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3403..3831
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3845..4174
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4241..5035
                     /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    5211..5332
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5381..5397)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5405..5421
                     /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(5429..5459)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5474..5495
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5783..6371)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6542..7402)
                     /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(7403..7507)
                     /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 ccaccatgcg cagactcagt
      721 tcctggagaa agatggcgac ggccgagaag cagaagcacg acgggcgggt gaagatcggc
      781 cactacatcc tgggggacac gcttggtgtc ggcaccttcg ggaaagtgaa ggtgggcaag
      841 cacgagttga ccggacataa agtggctgtg aagatactca accggcagaa gattcggagc
      901 cttgacgtgg tgggaaaaat ccgccgggag attcagaacc tgaagctgtt caggcaccct
      961 cacatcatca aactgtacca ggtcatcagt acaccatctg atattttcat ggtgatggaa
     1021 tatgtctctg gaggagagct atttgattat atctgtaaaa atggaaggtt ggacgaaaag
     1081 gaaagccgcc gtctgttcca gcagatcctt tccggtgtgg attattgtca caggcatatg
     1141 gtggtccaca gagatttgaa acctgagaac gtcctgcttg atgcacacat gaatgcaaag
     1201 atagccgact ttggtctttc aaacatgatg tcagatggtg aatttttaag aacaagctgt
     1261 ggctcaccca attatgccgc accagaagtc atttcaggaa gattgtacgc aggccccgag
     1321 gtggacatct ggagcagcgg ggtcattctc tatgctttgc tgtgtggaac cctccctttt
     1381 gatgatgacc atgtgccaac tctttttaag aagatatgtg atgggatctt ttatacccct
     1441 cagtacttaa acccttcagt aatcagcctt ttgaaacata tgctgcaggt ggatcccatg
     1501 aagagggccg caataaaaga tatcagggaa cacgagtggt ttaaacagga ccttccgaag
     1561 tatctctttc ctgaggaccc atcttatagt tcaaccatga tcgatgacga agccttgaaa
     1621 gaagtgtgtg agaagttcga gtgttcggag gaggaggtcc tcagctgcct gtacaacaga
     1681 aaccaccagg acccactagc cgtcgcctac cacctcatca tagacaacag gagaataatg
     1741 aatgaagcca aagatttcta cctagcaacc agcccacctg actctttcct ggacgaccac
     1801 catttaactc ggcctcaccc tgaaagagta ccgttcttgg ttgccgaaac accacgggcc
     1861 cggcacaccc tggatgaatt aaacccacag aaatccaaac accaaggtgt acggaaggca
     1921 aaatggcatt tgggaattcg aagtcaaagc cgacccaatg atatcatggc agaagtttgt
     1981 agagcaatca agcagttgga ttatgaatgg aaggttgtaa acccctatta tttgcgtgta
     2041 cgaaggaaga atcctgtgac aagcacattt tccaaaatga gtctacagct ataccaagtg
     2101 gatagtagga cttacttgtt ggatttccgt agtattgatg atgagattac agaagccaaa
     2161 tcagggactg ctactccaca gagatcggga tccatcagca actatcgatc ttgccaaagg
     2221 agtgactctg atgccgaagc tcaaggaaag ccctcagacg tctcccttac ctcatctgtc
     2281 acctccctcg actcctcccc tgtcgacgta gctccaagac caggaagtca tacaatagaa
     2341 ttttttgaaa tgtgtgcaaa tctaattaaa attcttgcac agctcgagta cccatacgac
     2401 gtacctgatt atgcaactta tccttatgat gtacctgatt atgctagcta tccttatgat
     2461 gtcccagact acgcatagtg agggcccgat atctcgacaa tcaacctctg gattacaaaa
     2521 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2581 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2641 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2701 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2761 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2821 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     2881 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     2941 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3001 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3061 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     3121 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3181 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     3241 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     3301 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     3361 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3421 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3481 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3541 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     3601 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3661 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3721 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     3781 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3841 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     3901 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     3961 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4021 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4081 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4141 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4201 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4261 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4321 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4381 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4441 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4501 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4561 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4621 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4681 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4741 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4801 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4861 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     4921 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     4981 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5041 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5101 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5161 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5221 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5281 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5341 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5401 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5461 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5521 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5581 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5641 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5701 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5761 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5821 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     5881 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     5941 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     6001 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6061 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6121 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6181 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6241 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6301 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6361 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6421 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6481 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6541 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6601 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6661 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6721 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6781 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6841 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     6901 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     6961 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7021 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7081 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7141 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7201 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7261 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7321 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7381 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7441 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7501 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7561 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7621 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7681 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
