LOCUS       sequence-274597-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 7 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-7
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        349..552
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     503..523
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    575..655
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      657..666
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(663..665,666..668,669..1382)
                     /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(663..665,666..668,669..1379)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(708..729)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(969..988)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1316..1337
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1723..1742)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1788..1843
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1842..1861)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2204..2220)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2204..2220)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2217..2239)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2228..2244
                     /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(2252..2258,2259..2276,2277..2282))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2297..2318
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2377..2572
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2423..2558
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2578..2712
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2615..2634)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2669..2688
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2780..2797)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2951..3539)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3031..3050)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3710..4501,4502..4570))
                     /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     4333..4352
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4571..4675)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4706..4733,1..348)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 tcaattacgg ggtcattagt tcatagccca tatatggagt tccgcgttac ataacttacg
       61 gtaaatggcc cgcctggctg accgcccaac gacccccgcc cattgacgtc aataatgacg
      121 tatgttccca tagtaacgcc aatagggact ttccattgac gtcaatgggt ggagtattta
      181 cggtaaactg cccacttggc agtacatcaa gtgtatcata tgccaagtac gccccctatt
      241 gacgtcaatg acggtaaatg gcccgcctgg cattatgccc agtacatgac cttatgggac
      301 tttcctactt ggcagtacat ctacgtatta gtcatcgcta ttaccatggt gatgcggttt
      361 tggcagtaca tcaatgggcg tggatagcgg tttgactcac ggggatttcc aagtctccac
      421 cccattgacg tcaatgggag tttgttttgg caccaaaatc aacgggactt tccaaaatgt
      481 cgtaacaact ccgccccatt gacgcaaatg ggcggtaggc gtgtacggtg ggaggtctat
      541 ataagcagag ctggtttagt gaaccgtcag atccgctagc gctaccggac tcagatctcg
      601 agctcaagct tcgaattctg cagtcgacgg taccgcgggc ccgggatcca ccggtcgcca
      661 ccatggtgag caagggcgag gagctgttca ccggggtggt gcccatcctg gtcgagctgg
      721 acggcgacgt aaacggccac aagttcagcg tgtccggcga gggcgagggc gatgccacct
      781 acggcaagct gaccctgaag ttcatctgca ccaccggcaa gctgcccgtg ccctggccca
      841 ccctcgtgac caccctgacc tacggcgtgc agtgcttcag ccgctacccc gaccacatga
      901 agcagcacga cttcttcaag tccgccatgc ccgaaggcta cgtccaggag cgcaccatct
      961 tcttcaagga cgacggcaac tacaagaccc gcgccgaggt gaagttcgag ggcgacaccc
     1021 tggtgaaccg catcgagctg aagggcatcg acttcaagga ggacggcaac atcctggggc
     1081 acaagctgga gtacaactac aacagccaca acgtctatat catggccgac aagcagaaga
     1141 acggcatcaa ggtgaacttc aagatccgcc acaacatcga ggacggcagc gtgcagctcg
     1201 ccgaccacta ccagcagaac acccccatcg gcgacggccc cgtgctgctg cccgacaacc
     1261 actacctgag cacccagtcc gccctgagca aagaccccaa cgagaagcgc gatcacatgg
     1321 tcctgctgga gttcgtgacc gccgccggga tcactctcgg catggacgag ctgtacaagt
     1381 aaagcggccg cactcctcag gtgcatgcct ctcctccctc tctggcaggg ccccatcctg
     1441 ggcagagggg cctggggctg ggcccagagt ccagccgtcc agctgctcct ttcccagttt
     1501 gatttcaata aatctgtcca ctccccgttt ttgggggtga tcgttttaac agccaagggt
     1561 gcatccttca tggtctgggc ttgcgtctgt cttgggggac ttattcgtcc tggctctctt
     1621 tggtccttgc tctggtggga catggaggca agtgttgaag gctgcctatc agaaggtggt
     1681 ggctggtgtg gccaatgccc tggctcacaa ataccactga gatctttttc cctctgccaa
     1741 aaattatggg gacatcatga agccccttga gcatctgact tctggctaat aaaggaaatt
     1801 tattttcatt gcaatagtgt gttggaattt tttgtgtctc tcactcggaa ggacatatgg
     1861 gagggcaaat catttaaaac atcagaatga gtatttggtt tagagtttgg caacatatgc
     1921 ccatatgctg gctgccatga acaaaggttg gctataaaga ggtcatcagt atatgaaaca
     1981 gccccctgct gtccattcct tattccatag aaaagccttg acttgaggtt agattttttt
     2041 tatattttgt tttgtgttat ttttttcttt aacatcccta aaattttcct tacatgtttt
     2101 actagccaga tttttcctcc tctcctgact actcccagtc atagctgtcc ctcttctctt
     2161 atggagatcc ctcgacctgc agcccaagct tggcgtaatc atggtcatag ctgtttcctg
     2221 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     2281 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     2341 ctttccagtc gggaaacctg tcgtgccagc ggatccgcat ctcaattagt cagcaaccat
     2401 agtcccgccc ctaactccgc ccatcccgcc cctaactccg cccagttccg cccattctcc
     2461 gccccatggc tgactaattt tttttattta tgcagaggcc gaggccgcct cggcctctga
     2521 gctattccag aagtagtgag gaggcttttt tggaggccta ggcttttgca aaaagctaac
     2581 ttgtttattg cagcttataa tggttacaaa taaagcaata gcatcacaaa tttcacaaat
     2641 aaagcatttt tttcactgca ttctagttgt ggtttgtcca aactcatcaa tgtatcttat
     2701 catgtctgga tccgctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     2761 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     2821 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     2881 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     2941 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     3001 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     3061 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     3121 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     3181 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     3241 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     3301 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     3361 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     3421 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     3481 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     3541 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     3601 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     3661 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     3721 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     3781 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     3841 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     3901 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     3961 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     4021 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     4081 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     4141 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     4201 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     4261 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     4321 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     4381 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     4441 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     4501 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     4561 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     4621 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     4681 ttccccgaaa agtgccacct gggtcgacat tgattattga ctagtaatag taa
//
