LOCUS       sequence-274599-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 9 of the pCMV-GFP-CENPBpA-fixed-length 
            library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4733)
  AUTHORS   Wu X, Bartel DP
  TITLE     Widespread Influence of 3'-End Structures on Mammalian mRNA 
            Processing and Stability.
  JOURNAL   Cell. 2017 May 18;169(5):905-917.e11. doi: 
            10.1016/j.cell.2017.04.036.
  PUBMED    28525757
REFERENCE   2  (bases 1 to 4733)
  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"; doi: 
            "10.1016/j.cell.2017.04"; journalName: "Cell"; date: "2017-05-18- 
            18"; volume: "169"; issue: "5"; pages: "905-917.e11"
COMMENT     Sequence Label: pCMV-GFP-CENPBpA-fixed-length-clone-9
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    105..126
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(141..146,147..164,165..171)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    179..195
                     /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)."
     primer_bind     184..206
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     203..219
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     203..219
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     562..581
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    580..635
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     681..700
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1041..1754,1755..1757,1758..1760))
                     /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(1044..1754,1755..1757,1758..1760))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1086..1107)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1435..1454
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1694..1715
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1757..1766
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1768..1848
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(1871..2074)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(1900..1920)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2075..2450
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2481..2585
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2586..2654,2655..3446)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2804..2823)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3617..4205
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4106..4125
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4359..4376
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    4444..4578
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4468..4487)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4522..4541
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(join(4584..4733,1..46))
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      4598..4733
                     /label=SV40 ori
                     /note="SV40 origin of replication"
ORIGIN
        1 gggcggagtt aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac
       61 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
      121 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
      181 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
      241 ctgcaggtcg agggatctcc ataagagaag agggacagct atgactggga gtagtcagga
      301 gaggaggaaa aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa
      361 aaataacaca aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa
      421 taaggaatgg acagcagggg gctgtttcat atactgatga cctctttata gccaaccttt
      481 gttcatggca gccagcatat gggcatatgt tgccaaactc taaaccaaat actcattctg
      541 atgttttaaa tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca
      601 acacactatt gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg
      661 cttcatgatg tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc
      721 cagggcattg gccacaccag ccaccacctt ctgataggca gccttcaaca cttgcctcca
      781 tgtcccacca gagcaaggac caaagagagc caggacgaat aagtccccca agacagacgc
      841 aagcccagac catgaaggat gcacccttgg ctgttaaaac gttcaccccc acaaaagggg
      901 agtggacaga tttattgaaa tcaaactggg aaaggagcag ctggacggct ggactctggg
      961 cccagcccca ggcccctctg cccaggatgg ggccctgcca gagagggagg agaggcatgc
     1021 acctgaggag tgcggccgct ttacttgtac agctcgtcca tgccgagagt gatcccggcg
     1081 gcggtcacga actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg
     1141 gcggactggg tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg
     1201 gtgttctgct ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc
     1261 ttgaagttca ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg
     1321 ttgtagttgt actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc
     1381 ttcagctcga tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg
     1441 tagttgccgt cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg
     1501 gacttgaaga agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg
     1561 taggtcaggg tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg
     1621 aacttcaggg tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac
     1681 ttgtggccgt ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc
     1741 tcctcgccct tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac
     1801 tgcagaattc gaagcttgag ctcgagatct gagtccggta gcgctagcgg atctgacggt
     1861 tcactaaacc agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg
     1921 tcaatggggc ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca
     1981 aactcccatt gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc
     2041 cacgcccatt gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt
     2101 agatgtactg ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg
     2161 gccatttacc gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta
     2221 ctgccaagtg ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta
     2281 ttggcgttac tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg
     2341 gtcagccagg cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat
     2401 gaactaatga ccccgtaatt gattactatt actagtcaat aatcaatgtc gacccaggtg
     2461 gcacttttcg gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa
     2521 atatgtatcc gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga
     2581 agagtatgag tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc
     2641 ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg
     2701 gtgcacgagt gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc
     2761 gccccgaaga acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat
     2821 tatcccgtat tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg
     2881 acttggttga gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag
     2941 aattatgcag tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa
     3001 cgatcggagg accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc
     3061 gccttgatcg ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca
     3121 cgatgcctgt agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc
     3181 tagcttcccg gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc
     3241 tgcgctcggc ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg
     3301 ggtctcgcgg tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta
     3361 tctacacgac ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag
     3421 gtgcctcact gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga
     3481 ttgatttaaa acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc
     3541 tcatgaccaa aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa
     3601 agatcaaagg atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa
     3661 aaaaaccacc gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc
     3721 cgaaggtaac tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt
     3781 agttaggcca ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc
     3841 tgttaccagt ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac
     3901 gatagttacc ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca
     3961 gcttggagcg aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg
     4021 ccacgcttcc cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag
     4081 gagagcgcac gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt
     4141 ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat
     4201 ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc
     4261 acatgttctt tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt
     4321 gagctgatac cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag
     4381 cggaagagcg cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca
     4441 gcggatccag acatgataag atacattgat gagtttggac aaaccacaac tagaatgcag
     4501 tgaaaaaaat gctttatttg tgaaatttgt gatgctattg ctttatttgt aaccattata
     4561 agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac
     4621 tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat aaaaaaaatt
     4681 agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg gat
//
