LOCUS       sequence-449690-        5189 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5189)
  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 5189)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl 
            Acad Sci U S A. 1999 Dec 7. 96(25):14517-22."
COMMENT     Sequence Label: pBI-MCS-EGFP
FEATURES             Location/Qualifiers
     source          1..5189
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     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    join(1508..1526,1527..1549,1550..1568,1569..1591,1592..1610,
                     1611..1633,1634..1652,1653..1675,1676..1694,1695..1717,
                     1718..1736,1737..1759,1760..1778)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     primer_bind     1921..1937
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1921..1937
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     CDS             join(1945..1947,1948..1950,1951..2664)
                     /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             join(1945..1947,1948..1950,1951..2661)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1990..2011)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(2251..2270)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2598..2619
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     polyA_signal    2701..2835
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2725..2744)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2779..2798
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3198..3216)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3284..3388
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3389..3457,3458..4249)
                     /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(3607..3626)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4420..5008
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4909..4928
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5162..5179
                     /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 aactcgactt tcacttttct ctatcactga tagggagtgg
     1741 taaactcgac tttcactttt ctctatcact gatagggagt ggtaaactcg acctatataa
     1801 gcagagctcg tttagtgaac cgtcagatcg cctggagacg ccatccacgc tgttttgacc
     1861 tccatagaag acaccgggac cgatccagcc tccgcggccc cgaattcctg cagcggccgc
     1921 tctagaacta gtggatcccc gcggatggtg agcaagggcg aggagctgtt caccggggtg
     1981 gtgcccatcc tggtcgagct ggacggcgac gtaaacggcc acaagttcag cgtgtccggc
     2041 gagggcgagg gcgatgccac ctacggcaag ctgaccctga agttcatctg caccaccggc
     2101 aagctgcccg tgccctggcc caccctcgtg accaccctga cctacggcgt gcagtgcttc
     2161 agccgctacc ccgaccacat gaagcagcac gacttcttca agtccgccat gcccgaaggc
     2221 tacgtccagg agcgcaccat cttcttcaag gacgacggca actacaagac ccgcgccgag
     2281 gtgaagttcg agggcgacac cctggtgaac cgcatcgagc tgaagggcat cgacttcaag
     2341 gaggacggca acatcctggg gcacaagctg gagtacaact acaacagcca caacgtctat
     2401 atcatggccg acaagcagaa gaacggcatc aaggtgaact tcaagatccg ccacaacatc
     2461 gaggacggca gcgtgcagct cgccgaccac taccagcaga acacccccat cggcgacggc
     2521 cccgtgctgc tgcccgacaa ccactacctg agcacccagt ccgccctgag caaagacccc
     2581 aacgagaagc gcgatcacat ggtcctgctg gagttcgtga ccgccgccgg gatcactctc
     2641 ggcatggacg agctgtacaa gtaagaattc gatatcaagc ttgtcgacct agttctagag
     2701 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
     2761 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
     2821 tgcaataaac aagttaacaa caacaattgc attcatttta tgtttcaggt tcagggggag
     2881 gtgtgggagg ttttttaaag caagtaaaac ctctacaaat gtggtatggc tgattatgat
     2941 cctgcaagcc tcgtcgtcct ggccggacca cgctatctgt gcaaggtccc cggccccgga
     3001 cgcgcgctcc atgagcagag cgcccgccgc cgaggcgaag actcgggcgg cgccctgccc
     3061 gtcccaccag gtcaacaggc ggtaaccggc ctcttcatcg ggaatgcgcg cgaccttcag
     3121 catcgccggc atgtccccct ggcggacggg aagtatccag ctcgaccaag ctggccgcgt
     3181 cgagtgaaga cgaaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     3241 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     3301 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     3361 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     3421 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     3481 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     3541 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     3601 agttctgcta tgtggcgcgg tattatcccg tgttgacgcc gggcaagagc aactcggtcg
     3661 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     3721 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     3781 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     3841 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     3901 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     3961 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     4021 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     4081 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     4141 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     4201 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     4261 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     4321 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     4381 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     4441 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     4501 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     4561 tactgttctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     4621 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     4681 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     4741 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     4801 acagcgtgag ctatgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     4861 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     4921 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     4981 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     5041 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     5101 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     5161 cagcgagtca gtgagcgagg aagcggaag
//
