LOCUS       V022542                 7715 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V022542
VERSION     V022542
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..1624
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="enhanced YFP"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EYFP"
     CDS             1631..3229
                     /label="PRKAA1(NM_001355028)"
                     /note="PRKAA1(NM_001355028)"
                     /gene="PRKAA1"
     polyA_signal    3282..3506
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     rep_origin      3552..3980
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3994..4323
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4390..5184
                     /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    5358..5479
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5528..5544)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5552..5568
                     /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(5576..5606)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5621..5642
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5930..6515)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6686..7546)
                     /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(7547..7651)
                     /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 gtgagcaagg gcgaggagct gttcaccggg gtggtgccca tcctggtcga
      961 gctggacggc gacgtaaacg gccacaagtt cagcgtgtcc ggcgagggcg agggcgatgc
     1021 cacctacggc aagctgaccc tgaagttcat ctgcaccacc ggcaagctgc ccgtgccctg
     1081 gcccaccctc gtgaccacct tcggctacgg cctgcagtgc ttcgcccgct accccgacca
     1141 catgaagcag cacgacttct tcaagtccgc catgcccgaa ggctacgtcc aggagcgcac
     1201 catcttcttc aaggacgacg gcaactacaa gacccgcgcc gaggtgaagt tcgagggcga
     1261 caccctggtg aaccgcatcg agctgaaggg catcgacttc aaggaggacg gcaacatcct
     1321 ggggcacaag ctggagtaca actacaacag ccacaacgtc tatatcatgg ccgacaagca
     1381 gaagaacggc atcaaggtga acttcaagat ccgccacaac atcgaggacg gcagcgtgca
     1441 gctcgccgac cactaccagc agaacacccc catcggcgac ggccccgtgc tgctgcccga
     1501 caaccactac ctgagctacc agtccgccct gagcaaagac cccaacgaga agcgcgatca
     1561 catggtcctg ctggagttcg tgaccgccgc cgggatcact ctcggcatgg acgagctgta
     1621 caaggaattc atgcaggtga aaaatataag aaagctcccc tgccaaatta ttcaatcagt
     1681 tggcaaacat gaattgactg ggcataaagt agctgtgaag atactcaatc gacagaagat
     1741 tcggagcctt gatgtggtag gaaaaatccg cagagaaatt cagaacctca agcttttcag
     1801 gcatcctcat ataattaaac tgtaccaggt catcagtaca ccatctgata ttttcatggt
     1861 gatggaatat gtctcaggag gagagctatt tgattatatc tgtaagaatg gaaggctgga
     1921 tgaaaaagaa agtcggcgtc tgttccaaca gatcctttct ggtgtggatt attgtcacag
     1981 gcatatggtg gtccatagag atttgaaacc tgaaaatgtc ctgcttgatg cacacatgaa
     2041 tgcaaagata gctgattttg gtctttcaaa catgatgtca gatggtgaat ttttaagaac
     2101 aagttgtggc tcacccaact atgctgcacc agaagtaatt tcaggaagat tgtatgcagg
     2161 cccagaggta gatatatgga gcagtggggt tattctctat gctttattat gtggaaccct
     2221 tccatttgat gatgaccatg tgccaactct ttttaagaag atatgtgatg ggatcttcta
     2281 tacccctcaa tatttaaatc cttctgtgat tagccttttg aaacatatgc tgcaggtgga
     2341 tcccatgaag agggccacaa tcaaagatat cagggaacat gaatggttta aacaggacct
     2401 tccaaaatat ctctttcctg aggatccatc atatagttca accatgattg atgatgaagc
     2461 cttaaaagaa gtatgtgaaa agtttgagtg ctcagaagag gaagttctca gctgtcttta
     2521 caacagaaat caccaggatc ctttggcagt tgcctaccat ctcataatag ataacaggag
     2581 aataatgaat gaagccaaag atttctattt ggcgacaagc ccacctgatt cttttcttga
     2641 tgatcatcac ctgactcggc cccatcctga aagagtacca ttcttggttg ctgaaacacc
     2701 aagggcacgc catacccttg atgaattaaa tccacagaaa tccaaacacc aaggtgtaag
     2761 gaaagcaaaa tggcatttag gaattagaag tcaaagtcga ccaaatgata ttatggcaga
     2821 agtatgtaga gcaatcaaac aattggatta tgaatggaag gttgtaaacc catattattt
     2881 gcgtgtacga aggaagaatc ctgtgacaag cacttactcc aaaatgagtc tacagttata
     2941 ccaagtggat agtagaactt atctactgga tttccgtagt attgatgatg aaattacaga
     3001 agccaaatca gggactgcta ctccacagag atcgggatca gttagcaact atcgatcttg
     3061 ccaaaggagt gattcagatg ctgaggctca aggaaaatcc tcagaagttt ctcttacctc
     3121 atctgtgacc tcacttgact cttctcctgt tgacctaact ccaagacctg gaagtcacac
     3181 aatagaattt tttgagatgt gtgcaaatct aattaaaatt cttgcacaaa ccggttagct
     3241 cgagtctaga gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca
     3301 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     3361 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     3421 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     3481 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     3541 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     3601 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     3661 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     3721 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     3781 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     3841 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     3901 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     3961 acaaaaattt aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc
     4021 ccaggctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg
     4081 tgtggaaagt ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag
     4141 tcagcaacca tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc
     4201 gcccattctc cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc
     4261 tctgcctctg agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc
     4321 aaaaagctcc cgggagcttg tatatccatt ttcggatctg atcaagagac aggatgagga
     4381 tcgtttcgca tgattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     4441 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     4501 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     4561 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     4621 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     4681 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     4741 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     4801 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     4861 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     4921 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     4981 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     5041 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     5101 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     5161 cgccttcttg acgagttctt ctgagcggga ctctggggtt cgaaatgacc gaccaagcga
     5221 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     5281 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     5341 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     5401 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     5461 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     5521 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     5581 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     5641 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     5701 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     5761 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     5821 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     5881 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     5941 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     6001 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     6061 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     6121 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     6181 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     6241 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     6301 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     6361 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     6421 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtttt tttgtttgca
     6481 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     6541 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     6601 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     6661 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     6721 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     6781 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     6841 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     6901 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     6961 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     7021 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     7081 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     7141 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     7201 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     7261 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     7321 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     7381 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     7441 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     7501 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     7561 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     7621 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     7681 tcgacggatc gggagatctc ccgatcccct atggt
//
