LOCUS       sequence-313744-        7047 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 7047)
  AUTHORS   Rajagopalan H, Jallepalli PV, Rago C, Velculescu VE, Kinzler KW, 
            Vogelstein B, Lengauer C
  TITLE     Inactivation of hCDC4 can cause chromosomal instability.
  JOURNAL   Nature. 2004 Mar 4. 428(6978):77-81.
  PUBMED    14999283
REFERENCE   2  (bases 1 to 7047)
  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: "Nature. 
            2004 Mar 4. 428(6978):77-81."
COMMENT     Sequence Label: Cyclin E-pBI-EGFP
FEATURES             Location/Qualifiers
     source          1..7047
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..39
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     intron          2118..2690
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     primer_bind     complement(2826..2845)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    2891..2946
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(2945..2964)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(3344..3361)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3515..4103)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3595..3614)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4274..5065,5066..5134))
                     /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"
     promoter        complement(5135..5239)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     5307..5325
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     polyA_signal    5688..5822
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5725..5744)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5779..5798
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(join(5859..6572,6573..6575,6576..6578))
                     /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             complement(join(5862..6572,6573..6575,6576..6578))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(5904..5925)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     6253..6272
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     6512..6533
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(6586..6602)
                     /label=SK primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6586..6602)
                     /label=pBluescriptSK
                     /note="For pBluescript vector"
     protein_bind    join(6745..6763,6764..6786,6787..6805,6806..6828,6829..6847,
                     6848..6870,6871..6889,6890..6912,6913..6931,6932..6954,
                     6955..6973,6974..6996,6997..7015)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
ORIGIN
        1 ggtaggcgtg tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc
       61 gcctggagac gccatccacg ctgttttgac ctccatagaa gacaccggga ccgatccagc
      121 ctccgcggcc ccgaattcga gctcggtacc cggggatcct ctagtcagct gacgcgcctc
      181 gaggcatgcc tgcacccccc ccccgaggag cagccggcgc ggccgccagc gcggtgtagg
      241 gggcaggcgc ggatcccgcc accgccgcgc gctcggcccg ccgactcccg gcgccgccgc
      301 cgccactgcc gtcgccgccg ccgcctgccg ggactggagc gcgccgtccg ccgcggacaa
      361 gaccctggcc tcaggccgga gcagccccat catgccgagg gagcgcaggg agcgggatgc
      421 gaaggagcgg gacaccatga aggaggacgg cggcgcggag ttctcggctc gctccaggaa
      481 gaggaaggca aacgtgaccg tttttttgca ggatccagat gaagaaatgg ccaaaatcga
      541 caggacggcg agggaccagt gtgggagcca gccttgggac aataatgcag tctgtgcaga
      601 cccctgctcc ctgatcccca cacctgacaa agaagatgat gaccgggttt acccaaactc
      661 aacgtgcaag cctcggatta ttgcaccatc cagaggctcc ccgctgcctg tactgagctg
      721 ggcaaataga gaggaagtct ggaaaatcat gttaaacaag gaaaagacat acttaaggga
      781 tcagcacttt cttgagcaac accctcttct gcagccaaaa atgcgagcaa ttcttctgga
      841 ttggttaatg gaggtgtgtg aagtctataa acttcacagg gagacctttt acttggcaca
      901 agatttcttt gaccggtata tggcgacaca agaaaatgtt gtaaaaactc ttttacagct
      961 tattgggatt tcatctttat ttattgcagc caaacttgag gaaatctatc ctccaaagtt
     1021 gcaccagttt gcgtatgtga cagatggagc ttgttcagga gatgaaattc tcaccatgga
     1081 attaatgatt atgaaactgt tggatctctg tgtcctggat gttgactgcc ttgaatttcc
     1141 ttatggtata cttgctgctt cggccttgta tcatttctcg tcatctgaat tgatgcaaaa
     1201 ggtttcaggg tatcagtggt gcgacataga gaactgtgtc aagtggatgg ttccatttgc
     1261 catggttata agggagacgg ggagctcaaa actgaagcac ttcaagggcg tcgctgatga
     1321 agatgcacac aacatacaga cccacagaga cagcttggat ttgctggaca aagcccgagc
     1381 aaagaaagcc atgttgtctg aacaaaatag ggcttctcct ctccccagtg ggctcctcac
     1441 cccgccacag agcggtaaga agcagagcag cgggccggaa atggcgtgac caccccatcc
     1501 ttctccacca aagacagttg cgcgcctgct ccacgttctc ttctgtctgt tgcagcggag
     1561 gcgtgcgttt gcttttacag atatctgaat ggaagagtgt ttcttccaca acagaagtat
     1621 ttctgtggat ggcatcaaac agggcaaagt gttttttatt gaatgcttat aggttttttt
     1681 taaataagtg ggtcaagtac accagccacc tccagacacc agtgcgtgct cccgatgctg
     1741 ctatggaagg tgctacttga cctaagggac tcccacaaca acaaaagctt gaagctgtgg
     1801 agggccacgg tggcgtggct ctcctcgcag gtgttctggg ctccgttgta ccaagtggag
     1861 caggtggttg cgggcaagcg ttgtgcagag cccatagcca gctgggcagg gggctgccct
     1921 ctccacatta tcagttgaca gtgtacaatg cctttgatga actgttttgt aagtgctgct
     1981 atatctatcc attttttaat aaagataata ctgtttttga gacccgggag ctcgaattcg
     2041 aaagatctct taattaaggt cgacagttta aacgctagcg cggcctcgac gatatctcta
     2101 gagctgagaa cttcagggtg agtttgggga cccttgattg ttctttcttt ttcgctattg
     2161 taaaattcat gttatatgga gggggcaaag ttttcagggt gttgtttaga atgggaagat
     2221 gtcccttgta tcaccatgga ccctcatgat aattttgttt ctttcacttt ctactctgtt
     2281 gacaaccatt gtctcctctt attttctttt cattttctgt aactttttcg ttaaacttta
     2341 gcttgcattt gtaacgaatt tttaaattca cttttgttta tttgtcagat tgtaagtact
     2401 ttctctaatc actttttttt caaggcaatc agggtatatt atattgtact tcagcacagt
     2461 tttagagaac aattgttata attaaatgat aaggtagaat atttctgcat ataaattctg
     2521 gctggcgtgg aaatattctt attggtagaa acaactacac cctggtcatc atcctgcctt
     2581 tctctttatg gttacaatga tatacactgt ttgagatgag gataaaatac tctgagtcca
     2641 aaccgggccc ctctgctaac catgttcatg ccttcttctc tttcctacag ctcctgggca
     2701 acgtgctggt tgttgtgctg tctcatcatt ttggcaaaga attcactcct caggtgcagg
     2761 ctgcctatca gaaggtggtg gctggtgtgg ccaatgccct ggctcacaaa taccactgag
     2821 atcgatcttt ttccctctgc caaaaattat ggggacatca tgaagcccct tgagcatctg
     2881 acttctggct aataaaggaa atttattttc attgcaatag tgtgttggaa ttttttgtgt
     2941 ctctcactcg gaaggacata tgggagggca aatcatttaa aacatcagaa tgagtatttg
     3001 gtttagagtt tggcaacata tgcccatatg ctggctgcca tgaacaaagg ttggctataa
     3061 agaggtcatc agtatatgaa acagccccct gctgtccatt ccttattcca tagaaaagcc
     3121 ttgacttgag gttagatttt ttttatattt tgttttgtgt tatttttttc tttaacatcc
     3181 ctaaaatttt ccttacatgt tttactagcc agatttttcc tcctctcctg actactccca
     3241 gtcatagctg tccctcttct cttatggaga tccctcgact gcattaatga atcggccaac
     3301 gcgcggggag aggcggtttg cgtattgggc gctcttccgc ttcctcgctc actgactcgc
     3361 tgcgctcggt cgttcggctg cggcgagcgg tatcagctca ctcaaaggcg gtaatacggt
     3421 tatccacaga atcaggggat aacgcaggaa agaacatgtg agcaaaaggc cagcaaaagg
     3481 ccaggaaccg taaaaaggcc gcgttgctgg cgtttttcca taggctccgc ccccctgacg
     3541 agcatcacaa aaatcgacgc tcaagtcaga ggtggcgaaa cccgacagga ctataaagat
     3601 accaggcgtt tccccctgga agctccctcg tgcgctctcc tgttccgacc ctgccgctta
     3661 ccggatacct gtccgccttt ctcccttcgg gaagcgtggc gctttctcat agctcacgct
     3721 gtaggtatct cagttcggtg taggtcgttc gctccaagct gggctgtgtg cacgaacccc
     3781 ccgttcagcc cgaccgctgc gccttatccg gtaactatcg tcttgagtcc aacccggtaa
     3841 gacacgactt atcgccactg gcagcagcca ctggtaacag gattagcaga gcgaggtatg
     3901 taggcggtgc tacagagttc ttgaagtggt ggcctaacta cggctacact agaagaacag
     3961 tatttggtat ctgcgctctg ctgaagccag ttaccttcgg aaaaagagtt ggtagctctt
     4021 gatccggcaa acaaaccacc gctggtagcg gtggtttttt tgtttgcaag cagcagatta
     4081 cgcgcagaaa aaaaggatct caagaagatc ctttgatctt ttctacgggg tctgacgctc
     4141 agtggaacga aaactcacgt taagggattt tggtcatgag attatcaaaa aggatcttca
     4201 cctagatcct tttaaattaa aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa
     4261 cttggtctga cagttaccaa tgcttaatca gtgaggcacc tatctcagcg atctgtctat
     4321 ttcgttcatc catagttgcc tgactccccg tcgtgtagat aactacgata cgggagggct
     4381 taccatctgg ccccagtgct gcaatgatac cgcgagaccc acgctcaccg gctccagatt
     4441 tatcagcaat aaaccagcca gccggaaggg ccgagcgcag aagtggtcct gcaactttat
     4501 ccgcctccat ccagtctatt aattgttgcc gggaagctag agtaagtagt tcgccagtta
     4561 atagtttgcg caacgttgtt gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg
     4621 gtatggcttc attcagctcc ggttcccaac gatcaaggcg agttacatga tcccccatgt
     4681 tgtgcaaaaa agcggttagc tccttcggtc ctccgatcgt tgtcagaagt aagttggccg
     4741 cagtgttatc actcatggtt atggcagcac tgcataattc tcttactgtc atgccatccg
     4801 taagatgctt ttctgtgact ggtgagtact caaccaagtc attctgagaa tagtgtatgc
     4861 ggcgaccgag ttgctcttgc ccggcgtcaa cacgggataa taccgcgcca catagcagaa
     4921 ctttaaaagt gctcatcatt ggaaaacgtt cttcggggcg aaaactctca aggatcttac
     4981 cgctgttgag atccagttcg atgtaaccca ctcgtgcacc caactgatct tcagcatctt
     5041 ttactttcac cagcgtttct gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg
     5101 gaataagggc gacacggaaa tgttgaatac tcatactctt cctttttcaa tattattgaa
     5161 gcatttatca gggttattgt ctcatgagcg gatacatatt tgaatgtatt tagaaaaata
     5221 aacaaatagg ggttccgcgc acatttcccc gaaaagtgcc acctgacgtc taagaaacca
     5281 ttattatcat gacattaacc tataaaaata ggcgtatcac gaggcccttt cgtcttcact
     5341 cgacgcggcc agcttggtcg agctggatac ttcccgtccg ccagggggac atgccggcga
     5401 tgctgaaggt cgcgcgcatt cccgatgaag aggccggtta ccgcctgttg acctggtggg
     5461 acgggcaggg cgccgcccga gtcttcgcct cggcggcggg cgctctgctc atggagcgcg
     5521 cgtccggggc cggggacctt gcacagatag cgtggtccgg ccaggacgac gaggcttgca
     5581 ggatcataat cagccatacc acatttgtag aggttttact tgctttaaaa aacctcccac
     5641 acctccccct gaacctgaaa cataaaatga atgcaattgt tgttgttaac ttgtttattg
     5701 cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt
     5761 tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga
     5821 tcctctagaa ctaggtcgac aagcttgata tcgaattctt acttgtacag ctcgtccatg
     5881 ccgagagtga tcccggcggc ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg
     5941 ttggggtctt tgctcagggc ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg
     6001 gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc
     6061 tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt tcttctgctt gtcggccatg
     6121 atatagacgt tgtggctgtt gtagttgtac tccagcttgt gccccaggat gttgccgtcc
     6181 tccttgaagt cgatgccctt cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc
     6241 acctcggcgc gggtcttgta gttgccgtcg tccttgaaga agatggtgcg ctcctggacg
     6301 tagccttcgg gcatggcgga cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg
     6361 ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc
     6421 ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt aggtggcatc gccctcgccc
     6481 tcgccggaca cgctgaactt gtggccgttt acgtcgccgt ccagctcgac caggatgggc
     6541 accaccccgg tgaacagctc ctcgcccttg ctcaccatcc gcggggatcc actagttcta
     6601 gagcggccgc tgcaggaatt cggggccgcg gaggctggat cggtcccggt gtcttctatg
     6661 gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta aacgagctct
     6721 gcttatatag gtcgagttta ccactcccta tcagtgatag agaaaagtga aagtcgagtt
     6781 taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc cctatcagtg
     6841 atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa aagtgaaagt
     6901 cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta ccactcccta
     6961 tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat agagaaaagt
     7021 gaaagtcgag ctcggtaccc gggtcga
//
