LOCUS       sequence-274603-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 13 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-13
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        330..533
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     484..504
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    556..636
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      638..647
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(644..646,647..649,650..1363)
                     /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(644..646,647..649,650..1360)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(689..710)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(950..969)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1297..1318
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1704..1723)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1769..1824
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1823..1842)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2185..2201)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2185..2201)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2198..2220)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2209..2225
                     /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(2233..2239,2240..2257,2258..2263))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2278..2299
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2358..2553
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2404..2539
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2559..2693
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2596..2615)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2650..2669
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2761..2778)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2932..3520)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3012..3031)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3691..4482,4483..4551))
                     /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     4314..4333
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4552..4656)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4687..4733,1..329)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 ttcatagccc atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct
       61 gaccgcccaa cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc
      121 caatagggac tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg
      181 cagtacatca agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat
      241 ggcccgcctg gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca
      301 tctacgtatt agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc
      361 gtggatagcg gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga
      421 gtttgttttg gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat
      481 tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag
      541 tgaaccgtca gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct
      601 gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga
      661 ggagctgttc accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca
      721 caagttcagc gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa
      781 gttcatctgc accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac
      841 ctacggcgtg cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa
      901 gtccgccatg cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa
      961 ctacaagacc cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct
     1021 gaagggcatc gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta
     1081 caacagccac aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt
     1141 caagatccgc cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa
     1201 cacccccatc ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc
     1261 cgccctgagc aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac
     1321 cgccgccggg atcactctcg gcatggacga gctgtacaag taaagcggcc gcactcctca
     1381 ggtgcatgcc tctcctccct ctctggcagg gccccatcct gggcagaggg gcctggggct
     1441 gggcccagag tccagccgtc cagctgctcc tttcccagtt tgatttcaat aaatctgtcc
     1501 actccccttt tgtggggctg aacgttttaa cagccaaggg tgcatccttc atggtctggg
     1561 cttgcgtctg tcttggggga cttattcgtc ctggctctct ttggtccttg ctctggtggg
     1621 acatggaggc aagtgttgaa ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc
     1681 ctggctcaca aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg
     1741 aagccccttg agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg
     1801 tgttggaatt ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa
     1861 catcagaatg agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg
     1921 aacaaaggtt ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc
     1981 ttattccata gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta
     2041 tttttttctt taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc
     2101 ctctcctgac tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg
     2161 cagcccaagc ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct
     2221 cacaattcca cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg
     2281 agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct
     2341 gtcgtgccag cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg
     2401 cccatcccgc ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt
     2461 ttttttattt atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga
     2521 ggaggctttt ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata
     2581 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     2641 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca
     2701 ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc
     2761 ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc
     2821 aaaggcggta atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc
     2881 aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag
     2941 gctccgcccc cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc
     3001 gacaggacta taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt
     3061 tccgaccctg ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct
     3121 ttctcatagc tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg
     3181 ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct
     3241 tgagtccaac ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat
     3301 tagcagagcg aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg
     3361 ctacactaga agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa
     3421 aagagttggt agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt
     3481 ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc
     3541 tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt
     3601 atcaaaaagg atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta
     3661 aagtatatat gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat
     3721 ctcagcgatc tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac
     3781 tacgatacgg gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg
     3841 ctcaccggct ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag
     3901 tggtcctgca actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt
     3961 aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt
     4021 gtcacgctcg tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt
     4081 tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt
     4141 cagaagtaag ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct
     4201 tactgtcatg ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt
     4261 ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac
     4321 cgcgccacat agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa
     4381 actctcaagg atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa
     4441 ctgatcttca gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca
     4501 aaatgccgca aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct
     4561 ttttcaatat tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga
     4621 atgtatttag aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc
     4681 tgggtcgaca ttgattattg actagtaata gtaatcaatt acggggtcat tag
//
