LOCUS       sequence-274604-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 14 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-14
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        386..490
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(491..559,560..1351)
                     /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(709..728)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1522..2110
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2011..2030
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2264..2281
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    2349..2483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2373..2392)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2427..2446
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(2489..2684)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2503..2638
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    2743..2764
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2779..2784,2785..2802,2803..2809)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2817..2833
                     /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     2822..2844
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2841..2857
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2841..2857
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     3200..3219
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    3218..3273
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     3319..3338
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(3679..4392,4393..4395,4396..4398))
                     /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(3682..4392,4393..4395,4396..4398))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(3724..3745)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     4073..4092
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     4332..4353
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      4395..4404
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    4406..4486
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(4509..4712)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(4538..4558)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        join(4713..4733,1..355)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tacgtagatg tactgccaag taggaaagtc ccataaggtc atgtactggg cataatgcca
       61 ggcgggccat ttaccgtcat tgacgtcaat agggggcgta cttggcatat gatacacttg
      121 atgtactgcc aagtgggcag tttaccgtaa atactccacc cattgacgtc aatggaaagt
      181 ccctattggc gttactatgg gaacatacgt cattattgac gtcaatgggc gggggtcgtt
      241 gggcggtcag ccaggcgggc catttaccgt aagttatgta acgcggaact ccatatatgg
      301 gctatgaact aatgaccccg taattgatta ctattactag tcaataatca atgtcgaccc
      361 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
      421 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
      481 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
      541 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
      601 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
      661 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
      721 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
      781 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
      841 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
      901 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
      961 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     1021 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     1081 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     1141 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     1201 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     1261 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     1321 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     1381 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     1441 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     1501 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     1561 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     1621 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgttc ttctagtgta
     1681 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     1741 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     1801 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     1861 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     1921 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     1981 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     2041 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     2101 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     2161 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     2221 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     2281 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     2341 atgcagcgga tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa
     2401 tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca
     2461 ttataagctg caataaacaa gttagctttt tgcaaaagcc taggcctcca aaaaagcctc
     2521 ctcactactt ctggaatagc tcagaggccg aggcggcctc ggcctctgca taaataaaaa
     2581 aaattagtca gccatggggc ggagaatggg cggaactggg cggagttagg ggcgggatgg
     2641 gcggagttag gggcgggact atggttgctg actaattgag atgcggatcc gctggcacga
     2701 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     2761 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     2821 gagcggataa caatttcaca caggaaacag ctatgaccat gattacgcca agcttgggct
     2881 gcaggtcgag ggatctccat aagagaagag ggacagctat gactgggagt agtcaggaga
     2941 ggaggaaaaa tctggctagt aaaacatgta aggaaaattt tagggatgtt aaagaaaaaa
     3001 ataacacaaa acaaaatata aaaaaaatct aacctcaagt caaggctttt ctatggaata
     3061 aggaatggac agcagggggc tgtttcatat actgatgacc tctttatagc caacctttgt
     3121 tcatggcagc cagcatatgg gcatatgttg ccaaactcta aaccaaatac tcattctgat
     3181 gttttaaatg atttgccctc ccatatgtcc ttccgagtga gagacacaaa aaattccaac
     3241 acactattgc aatgaaaata aatttccttt attagccaga agtcagatgc tcaaggggct
     3301 tcatgatgtc cccataattt ttggcagagg gaaaaagatc tcagtggtat ttgtgagcca
     3361 gggcattggc cacaccagcc accaccttct gataggcagc cttcaacact tgcctccatg
     3421 tcccaccaga gcaaggacca aagagagcca ggacgaataa gtcccccaag acagacgcaa
     3481 gcccagacca tgaaggatgc acccttggct gttaaaatgt tcacccccac aaaaggggag
     3541 tggacagatt tattgaaatc aaactgggaa aggagcagct ggacggctgg actctgggcc
     3601 cagccccagg cccctctgcc caggatgggg ccctgccaga gagggaggag aggcatgcac
     3661 ctgaggagtg cggccgcttt acttgtacag ctcgtccatg ccgagagtga tcccggcggc
     3721 ggtcacgaac tccagcagga ccatgtgatc gcgcttctcg ttggggtctt tgctcagggc
     3781 ggactgggtg ctcaggtagt ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt
     3841 gttctgctgg tagtggtcgg cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt
     3901 gaagttcacc ttgatgccgt tcttctgctt gtcggccatg atatagacgt tgtggctgtt
     3961 gtagttgtac tccagcttgt gccccaggat gttgccgtcc tccttgaagt cgatgccctt
     4021 cagctcgatg cggttcacca gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta
     4081 gttgccgtcg tccttgaaga agatggtgcg ctcctggacg tagccttcgg gcatggcgga
     4141 cttgaagaag tcgtgctgct tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta
     4201 ggtcagggtg gtcacgaggg tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa
     4261 cttcagggtc agcttgccgt aggtggcatc gccctcgccc tcgccggaca cgctgaactt
     4321 gtggccgttt acgtcgccgt ccagctcgac caggatgggc accaccccgg tgaacagctc
     4381 ctcgcccttg ctcaccatgg tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg
     4441 cagaattcga agcttgagct cgagatctga gtccggtagc gctagcggat ctgacggttc
     4501 actaaaccag ctctgcttat atagacctcc caccgtacac gcctaccgcc catttgcgtc
     4561 aatggggcgg agttgttacg acattttgga aagtcccgtt gattttggtg ccaaaacaaa
     4621 ctcccattga cgtcaatggg gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca
     4681 cgcccattga tgtactgcca aaaccgcatc accatggtaa tagcgatgac taa
//
