LOCUS       sequence-274592-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 2 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-2
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        329..532
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     483..503
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    555..635
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      637..646
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(643..645,646..648,649..1362)
                     /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(643..645,646..648,649..1359)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(688..709)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(949..968)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1296..1317
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1703..1722)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1768..1823
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1822..1841)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2184..2200)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2184..2200)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2197..2219)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2208..2224
                     /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(2232..2238,2239..2256,2257..2262))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2277..2298
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2357..2552
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2403..2538
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2558..2692
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2595..2614)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2649..2668
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2760..2777)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2931..3519)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3011..3030)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3690..4481,4482..4550))
                     /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     4313..4332
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4551..4655)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4686..4733,1..328)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcatagccca tatatggagt tccgcgttac ataacttacg gtaaatggcc cgcctggctg
       61 accgcccaac gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc
      121 aatagggact ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc
      181 agtacatcaa gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg
      241 gcccgcctgg cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat
      301 ctacgtatta gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg
      361 tggatagcgg tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag
      421 tttgttttgg caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt
      481 gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt
      541 gaaccgtcag atccgctagc gctaccggac tcagatctcg agctcaagct tcgaattctg
      601 cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca ccatggtgag caagggcgag
      661 gagctgttca ccggggtggt gcccatcctg gtcgagctgg acggcgacgt aaacggccac
      721 aagttcagcg tgtccggcga gggcgagggc gatgccacct acggcaagct gaccctgaag
      781 ttcatctgca ccaccggcaa gctgcccgtg ccctggccca ccctcgtgac caccctgacc
      841 tacggcgtgc agtgcttcag ccgctacccc gaccacatga agcagcacga cttcttcaag
      901 tccgccatgc ccgaaggcta cgtccaggag cgcaccatct tcttcaagga cgacggcaac
      961 tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc tggtgaaccg catcgagctg
     1021 aagggcatcg acttcaagga ggacggcaac atcctggggc acaagctgga gtacaactac
     1081 aacagccaca acgtctatat catggccgac aagcagaaga acggcatcaa ggtgaacttc
     1141 aagatccgcc acaacatcga ggacggcagc gtgcagctcg ccgaccacta ccagcagaac
     1201 acccccatcg gcgacggccc cgtgctgctg cccgacaacc actacctgag cacccagtcc
     1261 gccctgagca aagaccccaa cgagaagcgc gatcacatgg tcctgctgga gttcgtgacc
     1321 gccgccggga tcactctcgg catggacgag ctgtacaagt aaagcggccg cactcctcag
     1381 gtgcatgcct ctcctccctc tctggcaggg ccccatcctg ggcagagggg cctggggctg
     1441 ggcccagagt ccagccgtcc agctgctcct ttcccagttt gatttcaata aatctgtcca
     1501 ctcccctttt gtgggggtga acgttttaac agccaagggt gcatccttca tggtctgggc
     1561 ttgcgtctgt cttgggggac ttattcgtcc tggctctctt tggtccttgc tctggtggga
     1621 catggaggca agtgttgaag gctgcctatc agaaggtggt ggctggtgtg gccaatgccc
     1681 tggctcacaa ataccactga gatctttttc cctctgccaa aaattatggg gacatcatga
     1741 agccccttga gcatctgact tctggctaat aaaggaaatt tattttcatt gcaatagtgt
     1801 gttggaattt tttgtgtctc tcactcggaa ggacatatgg gagggcaaat catttaaaac
     1861 atcagaatga gtatttggtt tagagtttgg caacatatgc ccatatgctg gctgccatga
     1921 acaaaggttg gctataaaga ggtcatcagt atatgaaaca gccccctgct gtccattcct
     1981 tattccatag aaaagccttg acttgaggtt agattttttt tatattttgt tttgtgttat
     2041 ttttttcttt aacatcccta aaattttcct tacatgtttt actagccaga tttttcctcc
     2101 tctcctgact actcccagtc atagctgtcc ctcttctctt atggagatcc ctcgacctgc
     2161 agcccaagct tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc
     2221 acaattccac acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga
     2281 gtgagctaac tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg
     2341 tcgtgccagc ggatccgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     2401 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     2461 tttttattta tgcagaggcc gaggccgcct cggcctctga gctattccag aagtagtgag
     2521 gaggcttttt tggaggccta ggcttttgca aaaagctaac ttgtttattg cagcttataa
     2581 tggttacaaa taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca
     2641 ttctagttgt ggtttgtcca aactcatcaa tgtatcttat catgtctgga tccgctgcat
     2701 taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta ttgggcgctc ttccgcttcc
     2761 tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc gagcggtatc agctcactca
     2821 aaggcggtaa tacggttatc cacagaatca ggggataacg caggaaagaa catgtgagca
     2881 aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg
     2941 ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg
     3001 acaggactat aaagatacca ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt
     3061 ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc cttcgggaag cgtggcgctt
     3121 tctcatagct cacgctgtag gtatctcagt tcggtgtagg tcgttcgctc caagctgggc
     3181 tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt
     3241 gagtccaacc cggtaagaca cgacttatcg ccactggcag cagccactgg taacaggatt
     3301 agcagagcga ggtatgtagg cggtgctaca gagttcttga agtggtggcc taactacggc
     3361 tacactagaa gaacagtatt tggtatctgc gctctgctga agccagttac cttcggaaaa
     3421 agagttggta gctcttgatc cggcaaacaa accaccgctg gtagcggtgg tttttttgtt
     3481 tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag aagatccttt gatcttttct
     3541 acggggtctg acgctcagtg gaacgaaaac tcacgttaag ggattttggt catgagatta
     3601 tcaaaaagga tcttcaccta gatcctttta aattaaaaat gaagttttaa atcaatctaa
     3661 agtatatatg agtaaacttg gtctgacagt taccaatgct taatcagtga ggcacctatc
     3721 tcagcgatct gtctatttcg ttcatccata gttgcctgac tccccgtcgt gtagataact
     3781 acgatacggg agggcttacc atctggcccc agtgctgcaa tgataccgcg agacccacgc
     3841 tcaccggctc cagatttatc agcaataaac cagccagccg gaagggccga gcgcagaagt
     3901 ggtcctgcaa ctttatccgc ctccatccag tctattaatt gttgccggga agctagagta
     3961 agtagttcgc cagttaatag tttgcgcaac gttgttgcca ttgctacagg catcgtggtg
     4021 tcacgctcgt cgtttggtat ggcttcattc agctccggtt cccaacgatc aaggcgagtt
     4081 acatgatccc ccatgttgtg caaaaaagcg gttagctcct tcggtcctcc gatcgttgtc
     4141 agaagtaagt tggccgcagt gttatcactc atggttatgg cagcactgca taattctctt
     4201 actgtcatgc catccgtaag atgcttttct gtgactggtg agtactcaac caagtcattc
     4261 tgagaatagt gtatgcggcg accgagttgc tcttgcccgg cgtcaatacg ggataatacc
     4321 gcgccacata gcagaacttt aaaagtgctc atcattggaa aacgttcttc ggggcgaaaa
     4381 ctctcaagga tcttaccgct gttgagatcc agttcgatgt aacccactcg tgcacccaac
     4441 tgatcttcag catcttttac tttcaccagc gtttctgggt gagcaaaaac aggaaggcaa
     4501 aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt gaatactcat actcttcctt
     4561 tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata catatttgaa
     4621 tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa agtgccacct
     4681 gggtcgacat tgattattga ctagtaatag taatcaatta cggggtcatt agt
//
