LOCUS       Exported                5036 bp ds-DNA     circular SYN 07-OCT-2019
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5036)
  AUTHORS   vfarsam
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Oct 9, 2019 from SnapGene 5.0.0
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5036
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
                     /note="color: #ffffff"
     primer_bind     49..65
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     promoter        75..93
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff"
     misc_feature    complement(119..362)
                     /label=pT4 LIR
                     /note="color: #ff9900; direction: LEFT"
     enhancer        369..748
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
                     /note="color: #a6acb3"
     promoter        749..952
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
                     /note="color: #ffffff"
     intron          1044..1176
                     /label=chimeric intron
                     /note="chimera between introns from human beta-globin and 
                     immunoglobulin heavy chain genes"
                     /note="color: #------"
     misc_feature    1181..1190
                     /label=Kozak sequence
                     /note="color: #a6acb3"
     CDS             1187..1906
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /note="color: #00ff00"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     polyA_signal    1913..1961
                     /label=synthetic polyadenylation signal
                     /note="color: #a6acb3"
     polyA_signal    1913..1918
                     /label=PolyA\signal
                     /note="vntifkey=""25"" - label=PolyA\signal - 1835..1840"
                     /note="color: #a6acb3"
     polyA_site      1932..1933
                     /label=PolyA\Site
                     /note="vntifkey=""26"" - label=PolyA\Site - 1854..1855"
                     /note="color: #a6acb3"
     misc_feature    1941..1962
                     /label=G/T\rich\DSE
                     /note="vntifkey=""21"" - label=G/T\rich\DSE - 1863..1884"
                     /note="color: #a6acb3"
     misc_feature    1999..2264
                     /label=pT4 RIR
                     /note="color: #ff9900; direction: RIGHT"
     promoter        complement(2297..2315)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     primer_bind     complement(2336..2352)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: LEFT"
     protein_bind    2360..2376
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
                     /note="color: #31849b"
     promoter        complement(2384..2414)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
                     /note="color: #ffffff; direction: LEFT"
     protein_bind    2429..2450
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
                     /note="color: #31849b"
     rep_origin      complement(2738..3326)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     CDS             complement(3497..4357)
                     /label=AmpR
                     /note="vntifkey=""4"" - label=AmpR - 3455..4315"
                     /note="color: #993366; direction: LEFT"
     promoter        complement(4358..4462)
                     /gene="bla"
                     /label=AmpR promoter
                     /note="color: #ffffff; direction: LEFT"
     rep_origin      4489..4944
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
                     /note="color: #ffff00"
ORIGIN
        1 caaggcgatt aagttgggta acgccagggt tttcccagtc acgacgttgt aaaacgacgg
       61 ccagtgagcg cgcgtaatac gactcactat agggcgaatt ggagctagcg tttaaaccta
      121 cagttgaagt cggaagttta catacactta agttggagtc attaaaactc gtttttcaac
      181 tactccacaa atttcttgtt aacaaacaat agttttggca agtcagttag gacatctact
      241 ttgtgcatga cacaagtcat ttttccaaca attgtttaca gacagattat ttcacttata
      301 attcactgta tcacaattcc agtgggtcag aagtgtacat acacgcgctt gactgtgcct
      361 ttaagcttgg cattgattat tgactagtta ttaatagtaa tcaattacgg ggtcattagt
      421 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
      481 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      541 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      601 agtacatcaa gtgtatcata tgccaagtcc gccccctatt gacgtcaatg acggtaaatg
      661 gcccgcctgg cattatgccc agtacatgac cttacgggac tttcctactt ggcagtacat
      721 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca ccaatgggcg
      781 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      841 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaataacc ccgccccgtt
      901 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag gtcgtttagt
      961 gaaccgtcag atcactagta gctttattgc ggtagtttat cacagttaaa ttgctaacgc
     1021 agtcagtgct cgactgatca caggtaagta tcaaggttac aagacaggtt taaggaggcc
     1081 aatagaaact gggcttgtcg agacagagaa gattcttgcg tttctgatag gcacctattg
     1141 gtcttactga catccacttt gcctttctct ccacagggcg gccgccatgg tgagcaaggg
     1201 cgaggagctg ttcaccgggg tggtgcccat cctggtcgag ctggacggcg acgtaaacgg
     1261 ccacaagttc agcgtgtccg gcgagggcga gggcgatgcc acctacggca agctgaccct
     1321 gaagttcatc tgcaccaccg gcaagctgcc cgtgccctgg cccaccctcg tgaccaccct
     1381 gacctacggc gtgcagtgct tcagccgcta ccccgaccac atgaagcagc acgacttctt
     1441 caagtccgcc atgcccgaag gctacgtcca ggagcgcacc atcttcttca aggacgacgg
     1501 caactacaag acccgcgccg aggtgaagtt cgagggcgac accctggtga accgcatcga
     1561 gctgaagggc atcgacttca aggaggacgg caacatcctg gggcacaagc tggagtacaa
     1621 ctacaacagc cacaacgtct atatcatggc cgacaagcag aagaacggca tcaaggtgaa
     1681 cttcaagatc cgccacaaca tcgaggacgg cagcgtgcag ctcgccgacc actaccagca
     1741 gaacaccccc atcggcgacg gccccgtgct gctgcccgac aaccactacc tgagcaccca
     1801 gtccgccctg agcaaagacc ccaacgagaa gcgcgatcac atggtcctgc tggagttcgt
     1861 gaccgccgcc gggatcactc tcggcatgga cgagctgtac aagtaagaat tcaataaaat
     1921 atctttattt tcattacatc tgtgtgttgg ttttttgtgt gtaattcgag catcttaccg
     1981 ccatttattc ccagatcttt aaacaattta aaggcaatgc taccaaatac taagcgcgtg
     2041 tatgtacact tctgacccac tgggaatgtg atgaaagaaa taaaagctga aatgaatcat
     2101 tctctctact attattctga tatttcacat tcttaaaata aagtggtgat cctaactgac
     2161 cttaagacag ggaatcttta ctcggattaa atgtcaggaa ttgtgaaaaa gtgagtttaa
     2221 atgtatttgg ctaaggtgta tgtaaacttc cgacttcaac tgtagtttaa acgggcccgg
     2281 tacccagctt ttgttccctt tagtgagggt taattgcgcg cttggcgtaa tcatggtcat
     2341 agctgtttcc tgtgtgaaat tgttatccgc tcacaattcc acacaacata cgagccggaa
     2401 gcataaagtg taaagcctgg ggtgcctaat gagtgagcta actcacatta attgcgttgc
     2461 gctcactgcc cgctttccag tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc
     2521 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2581 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2641 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2701 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     2761 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     2821 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     2881 ttaccggata cctgtccgcc tttctccctt cgggaaccgt ggcgctttct catagctcac
     2941 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3001 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3061 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3121 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagaa
     3181 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3241 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3301 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3361 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3421 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3481 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3541 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3601 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3661 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3721 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     3781 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     3841 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     3901 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     3961 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4021 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4081 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4141 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4201 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4261 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4321 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4381 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4441 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctgac gcgccctgta
     4501 gcggcgcatt aagcgcggcg ggagtggtgg ttacgcgcag cgtgaccgct acacttgcca
     4561 gcgccctagc gcccgctcct ttcgctttct tcccttcctt tctcgccacg ttcgccggct
     4621 ttccccgtca agctctaaat cgggggctcc ctttagggtt ccgatttagt gctttacggc
     4681 acctcgaccc caaaaaactt gattagggtg atggttcacg tagtgggcca tcgccctgat
     4741 agacggtttt tcgccctttg acgttggagt ccacgttctt taatagtgga ctcttgttcc
     4801 aaactggaac aacactcaac cctatctcgg tctattcttt tgatttataa gggattttgc
     4861 cgatttcggc ctattggtta aaaaatgagc tgatttaaca aaaatttaac gcgaatttta
     4921 acaaaatatt aacgcttaca atttccattc gccattcagg ctgcgcaact gttgggaagg
     4981 gcgatcggtg cgggcctctt cgctattacg ccagctggcg aaagggggat gtgctg
//
