LOCUS       sequence-274600-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 10 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-10
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        359..562
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     513..533
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    585..665
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      667..676
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(673..675,676..678,679..1392)
                     /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(673..675,676..678,679..1389)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(718..739)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(979..998)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1326..1347
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1733..1752)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1798..1853
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1852..1871)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2214..2230)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2214..2230)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2227..2249)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2238..2254
                     /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(2262..2268,2269..2286,2287..2292))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2307..2328
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2387..2582
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2433..2568
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2588..2722
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2625..2644)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2679..2698
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2790..2807)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2961..3549)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3041..3060)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3720..4511,4512..4580))
                     /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     4343..4362
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4581..4685)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4716..4733,1..358)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 gtaatagtaa tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac
       61 ataacttacg gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc
      121 aataatgacg tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt
      181 ggagtattta cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac
      241 gccccctatt gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac
      301 cttatgggac tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt
      361 gatgcggttt tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc
      421 aagtctccac cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt
      481 tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg
      541 ggaggtctat ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac
      601 tcagatctcg agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca
      661 ccggtcgcca ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg
      721 gtcgagctgg acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc
      781 gatgccacct acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg
      841 ccctggccca ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc
      901 gaccacatga agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag
      961 cgcaccatct tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag
     1021 ggcgacaccc tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac
     1081 atcctggggc acaagctgga gtacaactac aacagccaca acgtctatat catggccgac
     1141 aagcagaaga acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc
     1201 gtgcagctcg ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg
     1261 cccgacaacc actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc
     1321 gatcacatgg tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag
     1381 ctgtacaagt aaagcggccg cactcctcag gtgcatgcct ctcctccctc tctggcaggg
     1441 ccccatcctg ggcagagggg cctggggctg ggcccagagt ccagccgtcc agctgctcct
     1501 ttcccagttt gatttcaata aatctggcca ctcccctttt gtaggggtta acgttttaac
     1561 agccaagggt gcatccttca tggtctgggc ttgcgtctgt cttgggggac ttattcgtcc
     1621 tggctctctt tggtccttgc tctggtggga catggaggca agtgttgaag gctgcctatc
     1681 agaaggtggt ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc
     1741 cctctgccaa aaattatggg gacatcatga agccccttga gcatctgact tctggctaat
     1801 aaaggaaatt tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa
     1861 ggacatatgg gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg
     1921 caacatatgc ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt
     1981 atatgaaaca gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt
     2041 agattttttt tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct
     2101 tacatgtttt actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc
     2161 ctcttctctt atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag
     2221 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     2281 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     2341 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt
     2401 cagcaaccat agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg
     2461 cccattctcc gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct
     2521 cggcctctga gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca
     2581 aaaagctaac ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa
     2641 tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa
     2701 tgtatcttat catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc
     2761 ggtttgcgta ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt
     2821 cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca
     2881 ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa
     2941 aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat
     3001 cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc
     3061 cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc
     3121 gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt
     3181 tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac
     3241 cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg
     3301 ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca
     3361 gagttcttga agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc
     3421 gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa
     3481 accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa
     3541 ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac
     3601 tcacgttaag ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta
     3661 aattaaaaat gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt
     3721 taccaatgct taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata
     3781 gttgcctgac tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc
     3841 agtgctgcaa tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac
     3901 cagccagccg gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag
     3961 tctattaatt gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac
     4021 gttgttgcca ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc
     4081 agctccggtt cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg
     4141 gttagctcct tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc
     4201 atggttatgg cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct
     4261 gtgactggtg agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc
     4321 tcttgcccgg cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc
     4381 atcattggaa aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc
     4441 agttcgatgt aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc
     4501 gtttctgggt gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca
     4561 cggaaatgtt gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt
     4621 tattgtctca tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt
     4681 ccgcgcacat ttccccgaaa agtgccacct gggtcgacat tgattattga cta
//
