LOCUS       Exported                5105 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pBI-MCS-EGFP
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5105)
  AUTHORS   Yu J, Zhang L, Hwang PM, Rago C, Kinzler KW, Vogelstein B
  TITLE     Identification and classification of p53-regulated genes.
  JOURNAL   Proc Natl Acad Sci U S A. 1999 Dec 7. 96(25):14517-22.
  PUBMED    10588737
REFERENCE   2  (bases 1 to 5105)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5105
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     370..389
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    388..443
                     /label=-beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     489..508
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     intron          644..1216
                     /label=-beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     promoter        complement(1437..1475)
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     protein_bind    1508..1526
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1550..1568
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1592..1610
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1634..1652
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1676..1694
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     primer_bind     1837..1853
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1837..1853
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             1861..2580
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     CDS             1861..2577
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1906..1927)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2167..2186)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2514..2535
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2617..2751
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2641..2660)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2695..2714
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3114..3132)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3200..3304
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3305..4165
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(3523..3542)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4336..4924
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4825..4844
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5078..5095
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagtcgag
       61 ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga ggaggaaaaa
      121 tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa ataacacaaa
      181 acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata aggaatggac
      241 agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt tcatggcagc
      301 cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat gttttaaatg
      361 atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac acactattgc
      421 aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct tcatgatgtc
      481 cccataattt ttggcagagg gaaaaagatc gatctcagtg gtatttgtga gccagggcat
      541 tggccacacc agccaccacc ttctgatagg cagcctgcac ctgaggagtg aattctttgc
      601 caaaatgatg agacagcaca acaaccagca cgttgcccag gagctgtagg aaagagaaga
      661 aggcatgaac atggttagca gaggggcccg gtttggactc agagtatttt atcctcatct
      721 caaacagtgt atatcattgt aaccataaag agaaaggcag gatgatgacc agggtgtagt
      781 tgtttctacc aataagaata tttccacgcc agccagaatt tatatgcaga aatattctac
      841 cttatcattt aattataaca attgttctct aaaactgtgc tgaagtacaa tataatatac
      901 cctgattgcc ttgaaaaaaa agtgattaga gaaagtactt acaatctgac aaataaacaa
      961 aagtgaattt aaaaattcgt tacaaatgca agctaaagtt taacgaaaaa gttacagaaa
     1021 atgaaaagaa aataagagga gacaatggtt gtcaacagag tagaaagtga aagaaacaaa
     1081 attatcatga gggtccatgg tgatacaagg gacatcttcc cattctaaac aacaccctga
     1141 aaactttgcc ccctccatat aacatgaatt ttacaatagc gaaaaagaaa gaacaatcaa
     1201 gggtccccaa actcaccctg aagttctcag ctctagagat atcgtcgagg ccgcgctagc
     1261 gtttaaactg tcgaccttaa ttaagagatc tgctcgaggc gcgtcagctg actagaggat
     1321 ccccgggtac cgagctcgaa ttcggggccg cggaggctgg atcggtcccg gtgtcttcta
     1381 tggaggtcaa aacagcgtgg atggcgtctc caggcgatct gacggttcac taaacgagct
     1441 ctgcttatat aggcctccca ccgtacacgc ctacctcgac ccgggtaccg agctcgactt
     1501 tcacttttct ctatcactga tagggagtgg taaactcgac tttcactttt ctctatcact
     1561 gatagggagt ggtaaactcg actttcactt ttctctatca ctgataggga gtggtaaact
     1621 cgactttcac ttttctctat cactgatagg gagtggtaaa ctcgactttc acttttctct
     1681 atcactgata gggagtggta aactcgacct atataagcag agctcgttta gtgaaccgtc
     1741 agatcgcctg gagacgccat ccacgctgtt ttgacctcca tagaagacac cgggaccgat
     1801 ccagcctccg cggccccgaa ttcctgcagc ggccgctcta gaactagtgg atccccgcgg
     1861 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
     1921 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
     1981 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
     2041 ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc gctaccccga ccacatgaag
     2101 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
     2161 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
     2221 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
     2281 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
     2341 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
     2401 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
     2461 tacctgagca cccagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
     2521 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtaa
     2581 gaattcgata tcaagcttgt cgacctagtt ctagaggatc cagacatgat aagatacatt
     2641 gatgagtttg gacaaaccac aactagaatg cagtgaaaaa aatgctttat ttgtgaaatt
     2701 tgtgatgcta ttgctttatt tgtaaccatt ataagctgca ataaacaagt taacaacaac
     2761 aattgcattc attttatgtt tcaggttcag ggggaggtgt gggaggtttt ttaaagcaag
     2821 taaaacctct acaaatgtgg tatggctgat tatgatcctg caagcctcgt cgtcctggcc
     2881 ggaccacgct atctgtgcaa ggtccccggc cccggacgcg cgctccatga gcagagcgcc
     2941 cgccgccgag gcgaagactc gggcggcgcc ctgcccgtcc caccaggtca acaggcggta
     3001 accggcctct tcatcgggaa tgcgcgcgac cttcagcatc gccggcatgt ccccctggcg
     3061 gacgggaagt atccagctcg accaagctgg ccgcgtcgag tgaagacgaa agggcctcgt
     3121 gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga cgtcaggtgg
     3181 cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa tacattcaaa
     3241 tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt gaaaaaggaa
     3301 gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg cattttgcct
     3361 tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag atcagttggg
     3421 tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg agagttttcg
     3481 ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt
     3541 atcccgtgtt gacgccgggc aagagcaact cggtcgccgc atacactatt ctcagaatga
     3601 cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga cagtaagaga
     3661 attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac ttctgacaac
     3721 gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc atgtaactcg
     3781 ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc gtgacaccac
     3841 gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac tacttactct
     3901 agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag gaccacttct
     3961 gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg gtgagcgtgg
     4021 gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta tcgtagttat
     4081 ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg ctgagatagg
     4141 tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata tactttagat
     4201 tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt ttgataatct
     4261 catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc ccgtagaaaa
     4321 gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct tgcaaacaaa
     4381 aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa ctctttttcc
     4441 gaaggtaact ggcttcagca gagcgcagat accaaatact gttcttctag tgtagccgta
     4501 gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc tgctaatcct
     4561 gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg actcaagacg
     4621 atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca cacagcccag
     4681 cttggagcga acgacctaca ccgaactgag atacctacag cgtgagctat gagaaagcgc
     4741 cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg tcggaacagg
     4801 agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt
     4861 tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc ggagcctatg
     4921 gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc cttttgctca
     4981 catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg cctttgagtg
     5041 agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc
     5101 ggaag
//
