LOCUS       sequence-274595-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 5 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-5
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(34..229)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      48..183
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    288..309
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(324..329,330..347,348..354)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    362..378
                     /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     367..389
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     386..402
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     386..402
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     745..764
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    763..818
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     864..883
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1224..1937,1938..1940,1941..1943))
                     /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(1227..1937,1938..1940,1941..1943))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1269..1290)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1618..1637
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1877..1898
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      1940..1949
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    1951..2031
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2054..2257)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2083..2103)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2258..2633
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2664..2768
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2769..2837,2838..3629)
                     /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(2987..3006)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3800..4388
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4289..4308
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4542..4559
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    join(4627..4733,1..28)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4651..4670)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     4705..4724
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
ORIGIN
        1 aaccattata agctgcaata aacaagttag ctttttgcaa aagcctaggc ctccaaaaaa
       61 gcctcctcac tacttctgga atagctcaga ggccgaggcg gcctcggcct ctgcataaat
      121 aaaaaaaatt agtcagccat ggggcggaga atgggcggaa ctgggcggag ttaggggcgg
      181 gatgggcgga gttaggggcg ggactatggt tgctgactaa ttgagatgcg gatccgctgg
      241 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
      301 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
      361 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagctt
      421 gggctgcagg tcgagggatc tccataagag aagagggaca gctatgactg ggagtagtca
      481 ggagaggagg aaaaatctgg ctagtaaaac atgtaaggaa aattttaggg atgttaaaga
      541 aaaaaataac acaaaacaaa atataaaaaa aatctaacct caagtcaagg cttttctatg
      601 gaataaggaa tggacagcag ggggctgttt catatactga tgacctcttt atagccaacc
      661 tttgttcatg gcagccagca tatgggcata tgttgccaaa ctctaaacca aatactcatt
      721 ctgatgtttt aaatgatttg ccctcccata tgtccttccg agtgagagac acaaaaaatt
      781 ccaacacact attgcaatga aaataaattt cctttattag ccagaagtca gatgctcaag
      841 gggcttcatg atgtccccat aatttttggc agagggaaaa agatctcagt ggtatttgtg
      901 agccagggca ttggccacac cagccaccac cttctgatag gcagccttca acacttgcct
      961 ccatgtccca ccagagcaag gaccaaagag agccaggacg aataagtccc ccaagacaga
     1021 cgcaagccca gaccatgaag gatgcaccct tggctgttaa aatgttcacc cccacaaaaa
     1081 gggagtggac agatttattg aaatcaaact gggaaaggag cagctggacg gctggactct
     1141 gggcccagcc ccaggcccct ctgcccagga tggggccctg ccagagaggg aggagaggca
     1201 tgcacctgag gagtgcggcc gctttacttg tacagctcgt ccatgccgag agtgatcccg
     1261 gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg gtctttgctc
     1321 agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc gtcgccgatg
     1381 ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat gttgtggcgg
     1441 atcttgaagt tcaccttgat gccgttcttc tgcttgtcgg ccatgatata gacgttgtgg
     1501 ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt gaagtcgatg
     1561 cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc ggcgcgggtc
     1621 ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc ttcgggcatg
     1681 gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa gcactgcacg
     1741 ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc ggtggtgcag
     1801 atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc ggacacgctg
     1861 aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac cccggtgaac
     1921 agctcctcgc ccttgctcac catggtggcg accggtggat cccgggcccg cggtaccgtc
     1981 gactgcagaa ttcgaagctt gagctcgaga tctgagtccg gtagcgctag cggatctgac
     2041 ggttcactaa accagctctg cttatataga cctcccaccg tacacgccta ccgcccattt
     2101 gcgtcaatgg ggcggagttg ttacgacatt ttggaaagtc ccgttgattt tggtgccaaa
     2161 acaaactccc attgacgtca atggggtgga gacttggaaa tccccgtgag tcaaaccgct
     2221 atccacgccc attgatgtac tgccaaaacc gcatcaccat ggtaatagcg atgactaata
     2281 cgtagatgta ctgccaagta ggaaagtccc ataaggtcat gtactgggca taatgccagg
     2341 cgggccattt accgtcattg acgtcaatag ggggcgtact tggcatatga tacacttgat
     2401 gtactgccaa gtgggcagtt taccgtaaat actccaccca ttgacgtcaa tggaaagtcc
     2461 ctattggcgt tactatggga acatacgtca ttattgacgt caatgggcgg gggtcgttgg
     2521 gcggtcagcc aggcgggcca tttaccgtaa gttatgtaac gcggaactcc atatatgggc
     2581 tatgaactaa tgaccccgta attgattact attactagtc aataatcaat gtcgacccag
     2641 gtggcacttt tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt
     2701 caaatatgta tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa
     2761 ggaagagtat gagtattcaa catttccgtg tcgcccttat tccctttttt gcagcatttt
     2821 gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt
     2881 tgggtgcacg agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt
     2941 ttcgccccga agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg
     3001 tattatcccg tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga
     3061 atgacttggt tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa
     3121 gagaattatg cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga
     3181 caacgatcgg aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa
     3241 ctcgccttga tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca
     3301 ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta
     3361 ctctagcttc ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac
     3421 ttctgcgctc ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc
     3481 gtgggtctcg cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag
     3541 ttatctacac gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga
     3601 taggtgcctc actgattaag cattggtaac tgtcagacca agtttactca tatatacttt
     3661 agattgattt aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata
     3721 atctcatgac caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag
     3781 aaaagatcaa aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa
     3841 caaaaaaacc accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt
     3901 ttccgaaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     3961 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     4021 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     4081 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     4141 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     4201 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     4261 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     4321 ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc
     4381 tatggaaaaa cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg
     4441 ctcacatgtt ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg
     4501 agtgagctga taccgctcgc cgtagccgaa cgaccgagcg cagcgagtca gtgagcgagg
     4561 aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat
     4621 gcagcggatc cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg
     4681 cagtgaaaaa aatgctttat ttgtgaaatt tgtgatgcta ttgctttatt tgt
//
