LOCUS       sequence-274593-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 3 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-3
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     96..113
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    181..315
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(205..224)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     259..278
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(321..516)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      335..470
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    575..596
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(611..616,617..634,635..641)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    649..665
                     /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     654..676
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     673..689
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     673..689
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     1032..1051
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    1050..1105
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     1151..1170
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(1511..2224,2225..2227,2228..2230))
                     /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(1514..2224,2225..2227,2228..2230))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(1556..1577)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     1905..1924
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     2164..2185
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      2227..2236
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    2238..2318
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(2341..2544)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(2370..2390)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        2545..2920
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2951..3055
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3056..3124,3125..3916)
                     /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(3274..3293)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4087..4675
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4576..4595
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag
       61 ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg
      121 aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat taatgcagcg
      181 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagtga
      241 aaaaaatgct ttatttgtga aatttgtgat gctattgctt tatttgtaac cattataagc
      301 tgcaataaac aagttagctt tttgcaaaag cctaggcctc caaaaaagcc tcctcactac
      361 ttctggaata gctcagaggc cgaggcggcc tcggcctctg cataaataaa aaaaattagt
      421 cagccatggg gcggagaatg ggcggaactg ggcggagtta ggggcgggat gggcggagtt
      481 aggggcggga ctatggttgc tgactaattg agatgcggat ccgctggcac gacaggtttc
      541 ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg
      601 caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat
      661 aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg ctgcaggtcg
      721 agggatctcc ataagagaag agggacagct atgactggga gtagtcagga gaggaggaaa
      781 aatctggcta gtaaaacatg taaggaaaat tttagggatg ttaaagaaaa aaataacaca
      841 aaacaaaata taaaaaaaat ctaacctcaa gtcaaggctt ttctatggaa taaggaatgg
      901 acagcagggg gctgtttcat atactgatga cctctttata gccaaccttt gttcatggca
      961 gccagcatat gggcatatgt tgccaaactc taaaccaaat actcattctg atgttttaaa
     1021 tgatttgccc tcccatatgt ccttccgagt gagagacaca aaaaattcca acacactatt
     1081 gcaatgaaaa taaatttcct ttattagcca gaagtcagat gctcaagggg cttcatgatg
     1141 tccccataat ttttggcaga gggaaaaaga tctcagtggt atttgtgagc cagggcattg
     1201 gccacaccag ccaccacctt ctgataggca gccttcaaca cttgcctcca tgtcccacca
     1261 gagcaaggac caaagagagc caggacgaat aagtccccca agacagacgc aagcccagac
     1321 catgaaggat gcacccttgg ctgttaaaac gattagcccc ataaaagggg agtggacaga
     1381 tttattgaaa tcaaactggg aaaggagcag ctggacggct ggactctggg cccagcccca
     1441 ggcccctctg cccaggatgg ggccctgcca gagagggagg agaggcatgc acctgaggag
     1501 tgcggccgct ttacttgtac agctcgtcca tgccgagagt gatcccggcg gcggtcacga
     1561 actccagcag gaccatgtga tcgcgcttct cgttggggtc tttgctcagg gcggactggg
     1621 tgctcaggta gtggttgtcg ggcagcagca cggggccgtc gccgatgggg gtgttctgct
     1681 ggtagtggtc ggcgagctgc acgctgccgt cctcgatgtt gtggcggatc ttgaagttca
     1741 ccttgatgcc gttcttctgc ttgtcggcca tgatatagac gttgtggctg ttgtagttgt
     1801 actccagctt gtgccccagg atgttgccgt cctccttgaa gtcgatgccc ttcagctcga
     1861 tgcggttcac cagggtgtcg ccctcgaact tcacctcggc gcgggtcttg tagttgccgt
     1921 cgtccttgaa gaagatggtg cgctcctgga cgtagccttc gggcatggcg gacttgaaga
     1981 agtcgtgctg cttcatgtgg tcggggtagc ggctgaagca ctgcacgccg taggtcaggg
     2041 tggtcacgag ggtgggccag ggcacgggca gcttgccggt ggtgcagatg aacttcaggg
     2101 tcagcttgcc gtaggtggca tcgccctcgc cctcgccgga cacgctgaac ttgtggccgt
     2161 ttacgtcgcc gtccagctcg accaggatgg gcaccacccc ggtgaacagc tcctcgccct
     2221 tgctcaccat ggtggcgacc ggtggatccc gggcccgcgg taccgtcgac tgcagaattc
     2281 gaagcttgag ctcgagatct gagtccggta gcgctagcgg atctgacggt tcactaaacc
     2341 agctctgctt atatagacct cccaccgtac acgcctaccg cccatttgcg tcaatggggc
     2401 ggagttgtta cgacattttg gaaagtcccg ttgattttgg tgccaaaaca aactcccatt
     2461 gacgtcaatg gggtggagac ttggaaatcc ccgtgagtca aaccgctatc cacgcccatt
     2521 gatgtactgc caaaaccgca tcaccatggt aatagcgatg actaatacgt agatgtactg
     2581 ccaagtagga aagtcccata aggtcatgta ctgggcataa tgccaggcgg gccatttacc
     2641 gtcattgacg tcaatagggg gcgtacttgg catatgatac acttgatgta ctgccaagtg
     2701 ggcagtttac cgtaaatact ccacccattg acgtcaatgg aaagtcccta ttggcgttac
     2761 tatgggaaca tacgtcatta ttgacgtcaa tgggcggggg tcgttgggcg gtcagccagg
     2821 cgggccattt accgtaagtt atgtaacgcg gaactccata tatgggctat gaactaatga
     2881 ccccgtaatt gattactatt actagtcaat aatcaatgtc gacccaggtg gcacttttcg
     2941 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     3001 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     3061 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     3121 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     3181 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     3241 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     3301 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     3361 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     3421 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     3481 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     3541 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     3601 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     3661 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     3721 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     3781 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     3841 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     3901 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     3961 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     4021 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     4081 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     4141 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     4201 tggcttcagc agagcgcaga taccaaatac tgttcttcta gtgtagccgt agttaggcca
     4261 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     4321 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     4381 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4441 aacgacctac accgaactga gatacctaca gcgtgagcta tgagaaagcg ccacgcttcc
     4501 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4561 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4621 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4681 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc aca
//
