LOCUS       V021401                 7843 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V021401
VERSION     V021401
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             649..1365
                     /codon_start=1
                     /note="mammalian codon-optimized"
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /transl_table=1
                     /translation="M,V,SKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKL
                     TLKFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFK
                     DDGNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNG
                     IKVNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMV
                     LLEFVTAAGITLGMDELYK"
                     /label="EGFP"
     CDS             1399..2553
                     /label="CAMK1D(NM_153498)"
                     /note="CAMK1D(NM_153498)"
                     /gene="CAMK1D"
     misc_feature    2619..3207
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    3273..3321
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      3523..3951
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3965..4294
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             4361..5155
                     /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    5331..5452
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(5501..5517)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5525..5541
                     /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(5549..5579)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5594..5615
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5903..6491)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6662..7522)
                     /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(7523..7627)
                     /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 gcgccaccat ggtgagcaag
      661 ggcgaggagc tgttcaccgg ggtggtgccc atcctggtcg agctggacgg cgacgtaaac
      721 ggccacaagt tcagcgtgtc cggcgagggc gagggcgatg ccacctacgg caagctgacc
      781 ctgaagttca tctgcaccac cggcaagctg cccgtgccct ggcccaccct cgtgaccacc
      841 ctgacctacg gcgtgcagtg cttcagccgc taccccgacc acatgaagca gcacgacttc
      901 ttcaagtccg ccatgcccga aggctacgtc caggagcgca ccatcttctt caaggacgac
      961 ggcaactaca agacccgcgc cgaggtgaag ttcgagggcg acaccctggt gaaccgcatc
     1021 gagctgaagg gcatcgactt caaggaggac ggcaacatcc tggggcacaa gctggagtac
     1081 aactacaaca gccacaacgt ctatatcatg gccgacaagc agaagaacgg catcaaggtg
     1141 aacttcaaga tccgccacaa catcgaggac ggcagcgtgc agctcgccga ccactaccag
     1201 cagaacaccc ccatcggcga cggccccgtg ctgctgcccg acaaccacta cctgagcacc
     1261 cagtccgccc tgagcaaaga ccccaacgag aagcgcgatc acatggtcct gctggagttc
     1321 gtgaccgccg ccgggatcac tctcggcatg gacgagctgt acaagggagg agggggaagc
     1381 ggtaccgagc tcggatccat ggcccgggag aacggcgaga gcagctcctc ctggaaaaag
     1441 caagctgaag acatcaagaa gatcttcgag ttcaaagaga ccctcggaac cggggccttt
     1501 tccgaagtgg ttttagctga agagaaggca actggcaagc tctttgctgt gaagtgtatc
     1561 cctaagaagg cgctgaaggg caaggaaagc agcatagaga atgagatagc cgtcctgaga
     1621 aagattaagc atgaaaatat tgttgccctg gaagacattt atgaaagccc aaatcacctg
     1681 tacttggtca tgcagctggt gtccggtgga gagctgtttg accggatagt ggagaagggg
     1741 ttttatacag agaaggatgc cagcactctg atccgccaag tcttggacgc cgtgtactat
     1801 ctccacagaa tgggcatcgt ccacagagac ctcaagcccg aaaatctctt gtactacagt
     1861 caagatgagg agtccaaaat aatgatcagt gactttggat tgtcaaaaat ggagggcaaa
     1921 ggagatgtga tgtccactgc ctgtggaact ccaggctatg tcgctcctga agtcctcgcc
     1981 cagaaacctt acagcaaagc cgttgactgc tggtccatcg gagtgattgc ctacatcttg
     2041 ctctgcggct accctccttt ttatgatgaa aatgactcca agctctttga gcagatcctc
     2101 aaggcggaat atgagtttga ctctccctac tgggatgaca tctccgactc tgcaaaagac
     2161 ttcattcgga acctgatgga gaaggacccg aataaaagat acacgtgtga gcaggcagct
     2221 cggcacccat ggatcgctgg tgacacagcc ctcaacaaaa acatccacga gtccgtcagc
     2281 gcccagatcc ggaaaaactt tgccaagagc aaatggagac aagcatttaa tgccacggcc
     2341 gtcgtcagac atatgagaaa actacacctc ggcagcagcc tggacagttc aaatgcaagt
     2401 gtttcgagca gcctcagttt ggccagccaa aaagactgtc tggcaccttc cacgctctgt
     2461 agtttcattt cttcttcgtc gggggtctca ggagttggag ccgagcggag acccaggccc
     2521 accactgtga cggcagtgca ctctggaagc aagtgagaat tctgcagata tccagcacag
     2581 tggcggccgc tcgagtctag agggcccgat atctcgacaa tcaacctctg gattacaaaa
     2641 tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta tgtggatacg
     2701 ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt ttctcctcct
     2761 tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc aggcaacgtg
     2821 gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt gccaccacct
     2881 gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg gaactcatcg
     2941 ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac aattccgtgg
     3001 tgttgtcggg gaagctgacg tcctttccat ggctgctcgc ctgtgttgcc acctggattc
     3061 tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac cttccttccc
     3121 gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct cagacgagtc
     3181 ggatctccct ttgggccgcc tccccgcctg gaaacggggg aggctaactg aaacacggaa
     3241 ggagacaata ccggaaggaa cccgcgctat gacggcaata aaaagacaga ataaaacgca
     3301 cgggtgttgg gtcgtttgtt cataaacgcg gggttcggtc ccagggctgg cactctgtcg
     3361 ataccccacc gagaccccat tggggccaat acgcccgcgt ttcttccttt tccccacccc
     3421 accccccaag ttcgggtgaa ggcccagggc tcgcagccaa cgtcggggcg gcaggccctg
     3481 ccatagcaga tctgcgcagc tggggctcta gggggtatcc ccacgcgccc tgtagcggcg
     3541 cattaagcgc ggcgggtgtg gtggttacgc gcagcgtgac cgctacactt gccagcgccc
     3601 tagcgcccgc tcctttcgct ttcttccctt cctttctcgc cacgttcgcc ggctttcccc
     3661 gtcaagctct aaatcggggc atccctttag ggttccgatt tagtgcttta cggcacctcg
     3721 accccaaaaa acttgattag ggtgatggtt cacgtagtgg gccatcgccc tgatagacgg
     3781 tttttcgccc tttgacgttg gagtccacgt tctttaatag tggactcttg ttccaaactg
     3841 gaacaacact caaccctatc tcggtctatt cttttgattt ataagggatt ttggggattt
     3901 cggcctattg gttaaaaaat gagctgattt aacaaaaatt taacgcgaat taattctgtg
     3961 gaatgtgtgt cagttagggt gtggaaagtc cccaggctcc ccagcaggca gaagtatgca
     4021 aagcatgcat ctcaattagt cagcaaccag gtgtggaaag tccccaggct ccccagcagg
     4081 cagaagtatg caaagcatgc atctcaatta gtcagcaacc atagtcccgc ccctaactcc
     4141 gcccatcccg cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat
     4201 tttttttatt tatgcagagg ccgaggccgc ctctgcctct gagctattcc agaagtagtg
     4261 aggaggcttt tttggaggcc taggcttttg caaaaagctc ccgggagctt gtatatccat
     4321 tttcggatct gatcaagaga caggatgagg atcgtttcgc atgattgaac aagatggatt
     4381 gcacgcaggt tctccggccg cttgggtgga gaggctattc ggctatgact gggcacaaca
     4441 gacaatcggc tgctctgatg ccgccgtgtt ccggctgtca gcgcaggggc gcccggttct
     4501 ttttgtcaag accgacctgt ccggtgccct gaatgaactg caggacgagg cagcgcggct
     4561 atcgtggctg gccacgacgg gcgttccttg cgcagctgtg ctcgacgttg tcactgaagc
     4621 gggaagggac tggctgctat tgggcgaagt gccggggcag gatctcctgt catctcacct
     4681 tgctcctgcc gagaaagtat ccatcatggc tgatgcaatg cggcggctgc atacgcttga
     4741 tccggctacc tgcccattcg accaccaagc gaaacatcgc atcgagcgag cacgtactcg
     4801 gatggaagcc ggtcttgtcg atcaggatga tctggacgaa gagcatcagg ggctcgcgcc
     4861 agccgaactg ttcgccaggc tcaaggcgcg catgcccgac ggcgaggatc tcgtcgtgac
     4921 ccatggcgat gcctgcttgc cgaatatcat ggtggaaaat ggccgctttt ctggattcat
     4981 cgactgtggc cggctgggtg tggcggaccg ctatcaggac atagcgttgg ctacccgtga
     5041 tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc ctcgtgcttt acggtatcgc
     5101 cgctcccgat tcgcagcgca tcgccttcta tcgccttctt gacgagttct tctgagcggg
     5161 actctggggt tcgcgaaatg accgaccaag cgacgcccaa cctgccatca cgagatttcg
     5221 attccaccgc cgccttctat gaaaggttgg gcttcggaat cgttttccgg gacgccggct
     5281 ggatgatcct ccagcgcggg gatctcatgc tggagttctt cgcccacccc aacttgttta
     5341 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     5401 ttttttcact gcattctagt tgtggtttgt ccaaactcat caatgtatct tatcatgtct
     5461 gtataccgtc gacctctagc tagagcttgg cgtaatcatg gtcatagctg tttcctgtgt
     5521 gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata aagtgtaaag
     5581 cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca ctgcccgctt
     5641 tccagtcggg aaacctgtcg tgccagctgc attaatgaat cggccaacgc gcggggagag
     5701 gcggtttgcg tattgggcgc tcttccgctt cctcgctcac tgactcgctg cgctcggtcg
     5761 ttcggctgcg gcgagcggta tcagctcact caaaggcggt aatacggtta tccacagaat
     5821 caggggataa cgcaggaaag aacatgtgag caaaaggcca gcaaaaggcc aggaaccgta
     5881 aaaaggccgc gttgctggcg tttttccata ggctccgccc ccctgacgag catcacaaaa
     5941 atcgacgctc aagtcagagg tggcgaaacc cgacaggact ataaagatac caggcgtttc
     6001 cccctggaag ctccctcgtg cgctctcctg ttccgaccct gccgcttacc ggatacctgt
     6061 ccgcctttct cccttcggga agcgtggcgc tttctcaatg ctcacgctgt aggtatctca
     6121 gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca cgaacccccc gttcagcccg
     6181 accgctgcgc cttatccggt aactatcgtc ttgagtccaa cccggtaaga cacgacttat
     6241 cgccactggc agcagccact ggtaacagga ttagcagagc gaggtatgta ggcggtgcta
     6301 cagagttctt gaagtggtgg cctaactacg gctacactag aaggacagta tttggtatct
     6361 gcgctctgct gaagccagtt accttcggaa aaagagttgg tagctcttga tccggcaaac
     6421 aaaccaccgc tggtagcggt ggtttttttg tttgcaagca gcagattacg cgcagaaaaa
     6481 aaggatctca agaagatcct ttgatctttt ctacggggtc tgacgctcag tggaacgaaa
     6541 actcacgtta agggattttg gtcatgagat tatcaaaaag gatcttcacc tagatccttt
     6601 taaattaaaa atgaagtttt aaatcaatct aaagtatata tgagtaaact tggtctgaca
     6661 gttaccaatg cttaatcagt gaggcaccta tctcagcgat ctgtctattt cgttcatcca
     6721 tagttgcctg actccccgtc gtgtagataa ctacgatacg ggagggctta ccatctggcc
     6781 ccagtgctgc aatgataccg cgagacccac gctcaccggc tccagattta tcagcaataa
     6841 accagccagc cggaagggcc gagcgcagaa gtggtcctgc aactttatcc gcctccatcc
     6901 agtctattaa ttgttgccgg gaagctagag taagtagttc gccagttaat agtttgcgca
     6961 acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc gtcgtttggt atggcttcat
     7021 tcagctccgg ttcccaacga tcaaggcgag ttacatgatc ccccatgttg tgcaaaaaag
     7081 cggttagctc cttcggtcct ccgatcgttg tcagaagtaa gttggccgca gtgttatcac
     7141 tcatggttat ggcagcactg cataattctc ttactgtcat gccatccgta agatgctttt
     7201 ctgtgactgg tgagtactca accaagtcat tctgagaata gtgtatgcgg cgaccgagtt
     7261 gctcttgccc ggcgtcaata cgggataata ccgcgccaca tagcagaact ttaaaagtgc
     7321 tcatcattgg aaaacgttct tcggggcgaa aactctcaag gatcttaccg ctgttgagat
     7381 ccagttcgat gtaacccact cgtgcaccca actgatcttc agcatctttt actttcacca
     7441 gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc aaaaaaggga ataagggcga
     7501 cacggaaatg ttgaatactc atactcttcc tttttcaata ttattgaagc atttatcagg
     7561 gttattgtct catgagcgga tacatatttg aatgtattta gaaaaataaa caaatagggg
     7621 ttccgcgcac atttccccga aaagtgccac ctgacgtcga cggatcggga gatctcccga
     7681 tcccctatgg tcgactctca gtacaatctg ctctgatgcc gcatagttaa gccagtatct
     7741 gctccctgct tgtgtgttgg aggtcgctga gtagtgcgcg agcaaaattt aagctacaac
     7801 aaggcaaggc ttgaccgaca attgcatgaa gaatctgctt agg
//
