LOCUS       sequence-227855-        4728 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 11 of the pCMV-GFP-CENPBpA-fixed-length 
            library.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4728)
  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 4728)
  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-11
FEATURES             Location/Qualifiers
     source          1..4728
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..375
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        376..579
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     530..550
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    602..682
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      684..693
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(690..692,693..695,696..1409)
                     /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(690..692,693..695,696..1406)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(735..756)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(996..1015)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1343..1364
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1750..1769)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1815..1870
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1869..1888)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2231..2247)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2231..2247)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2244..2266)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2255..2271
                     /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)."
     promoter        complement(join(2279..2285,2286..2303,2304..2309))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2324..2345
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2404..2599
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2450..2585
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2605..2739
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2642..2661)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2696..2715
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2807..2824)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2978..3566)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3058..3077)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3737..4528,4529..4597))
                     /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     4360..4379
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4598..4702)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgata atagtaatca attacggggt cattagttca tagcccatat
       61 atggagttcc gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac
      121 ccccgcccat tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc
      181 cattgacgtc aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg
      241 tatcatatgc caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat
      301 tatgcccagt acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc
      361 atcgctatta ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt
      421 gactcacggg gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac
      481 caaaatcaac gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc
      541 ggtaggcgtg tacggtggga ggtctatata agcagagctg gtttagtgaa ccgtcagatc
      601 cgctagcgct accggactca gatctcgagc tcaagcttcg aattctgcag tcgacggtac
      661 cgcgggcccg ggatccaccg gtcgccacca tggtgagcaa gggcgaggag ctgttcaccg
      721 gggtggtgcc catcctggtc gagctggacg gcgacgtaaa cggccacaag ttcagcgtgt
      781 ccggcgaggg cgagggcgat gccacctacg gcaagctgac cctgaagttc atctgcacca
      841 ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac cctgacctac ggcgtgcagt
      901 gcttcagccg ctaccccgac cacatgaagc agcacgactt cttcaagtcc gccatgcccg
      961 aaggctacgt ccaggagcgc accatcttct tcaaggacga cggcaactac aagacccgcg
     1021 ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat cgagctgaag ggcatcgact
     1081 tcaaggagga cggcaacatc ctggggcaca agctggagta caactacaac agccacaacg
     1141 tctatatcat ggccgacaag cagaagaacg gcatcaaggt gaacttcaag atccgccaca
     1201 acatcgagga cggcagcgtg cagctcgccg accactacca gcagaacacc cccatcggcg
     1261 acggccccgt gctgctgccc gacaaccact acctgagcac ccagtccgcc ctgagcaaag
     1321 accccaacga gaagcgcgat cacatggtcc tgctggagtt cgtgaccgcc gccgggatca
     1381 ctctcggcat ggacgagctg tacaagtaaa gcggccgcac tcctcaggtg catgcctctc
     1441 ctccctctct ggcagggccc catcctgggc agaggggcct ggggctgggc ccagagtcca
     1501 gccgtccagc tgctcctttc ccagtttgat ttcaataaat ctgtccactc cccttttgtg
     1561 ggggtgaacg ttttaacagc caagggtgca tccttcatgg tctgggcttg cgtctgtctt
     1621 gggggactta ttcgtcctgg ctctctttgg tccttgctct ggtgggacat ggaggcaagt
     1681 gttgaaggct gcctatcaga aggtggtggc tggtgtggcc aatgccctgg ctcacaaata
     1741 ccactgagat ctttttccct ctgccaaaaa ttatggggac atcatgaagc cccttgagca
     1801 tctgacttct ggctaataaa ggaaatttat tttcattgca atagtgtgtt ggaatttttt
     1861 gtgtctctca ctcggaagga catatgggag ggcaaatcat ttaaaacatc agaatgagta
     1921 tttggtttag agtttggcaa catatgccca tatgctggct gccatgaaca aaggttggct
     1981 ataaagaggt catcagtata tgaaacagcc ccctgctgtc cattccttat tccatagaaa
     2041 agccttgact tgaggttaga ttttttttat attttgtttt gtgttatttt tttctttaac
     2101 atccctaaaa ttttccttac atgttttact agccagattt ttcctcctct cctgactact
     2161 cccagtcata gctgtccctc ttctcttatg gagatccctc gacctgcagc ccaagcttgg
     2221 cgtaatcatg gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca
     2281 acatacgagc cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca
     2341 cattaattgc gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga
     2401 tccgcatctc aattagtcag caaccatagt cccgccccta actccgccca tcccgcccct
     2461 aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt ttatttatgc
     2521 agaggccgag gccgcctcgg cctctgagct attccagaag tagtgaggag gcttttttgg
     2581 aggcctaggc ttttgcaaaa agctaacttg tttattgcag cttataatgg ttacaaataa
     2641 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     2701 ttgtccaaac tcatcaatgt atcttatcat gtctggatcc gctgcattaa tgaatcggcc
     2761 aacgcgcggg gagaggcggt ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact
     2821 cgctgcgctc ggtcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     2881 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     2941 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3001 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3061 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3121 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct caatgctcac
     3181 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3241 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3301 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3361 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     3421 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     3481 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     3541 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     3601 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gagattatca aaaaggatct
     3661 tcacctagat ccttttaaat taaaaatgaa gttttaaatc aatctaaagt atatatgagt
     3721 aaacttggtc tgacagttac caatgcttaa tcagtgaggc acctatctca gcgatctgtc
     3781 tatttcgttc atccatagtt gcctgactcc ccgtcgtgta gataactacg atacgggagg
     3841 gcttaccatc tggccccagt gctgcaatga taccgcgaga cccacgctca ccggctccag
     3901 atttatcagc aataaaccag ccagccggaa gggccgagcg cagaagtggt cctgcaactt
     3961 tatccgcctc catccagtct attaattgtt gccgggaagc tagagtaagt agttcgccag
     4021 ttaatagttt gcgcaacgtt gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt
     4081 ttggtatggc ttcattcagc tccggttccc aacgatcaag gcgagttaca tgatccccca
     4141 tgttgtgcaa aaaagcggtt agctccttcg gtcctccgat cgttgtcaga agtaagttgg
     4201 ccgcagtgtt atcactcatg gttatggcag cactgcataa ttctcttact gtcatgccat
     4261 ccgtaagatg cttttctgtg actggtgagt actcaaccaa gtcattctga gaatagtgta
     4321 tgcggcgacc gagttgctct tgcccggcgt caatacggga taataccgcg ccacatagca
     4381 gaactttaaa agtgctcatc attggaaaac gttcttcggg gcgaaaactc tcaaggatct
     4441 taccgctgtt gagatccagt tcgatgtaac ccactcgtgc acccaactga tcttcagcat
     4501 cttttacttt caccagcgtt tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa
     4561 agggaataag ggcgacacgg aaatgttgaa tactcatact cttccttttt caatattatt
     4621 gaagcattta tcagggttat tgtctcatga gcggatacat atttgaatgt atttagaaaa
     4681 ataaacaaat aggggttccg cgcacatttc cccgaaaagt gccacctg
//
