LOCUS       sequence-274602-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 12 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-12
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        350..553
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     504..524
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    576..656
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      658..667
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(664..666,667..669,670..1383)
                     /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(664..666,667..669,670..1380)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(709..730)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(970..989)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1317..1338
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1724..1743)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1789..1844
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1843..1862)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2205..2221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2205..2221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2218..2240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2229..2245
                     /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(2253..2259,2260..2277,2278..2283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2298..2319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2378..2573
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2424..2559
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2579..2713
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2616..2635)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2670..2689
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2781..2798)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2952..3540)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3032..3051)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3711..4502,4503..4571))
                     /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     4334..4353
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4572..4676)
                     /gene="bla"
                     /label=AmpR promoter
     enhancer        join(4707..4733,1..349)
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
ORIGIN
        1 atcaattacg gggtcattag ttcatagccc atatatggag ttccgcgtta cataacttac
       61 ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc ccattgacgt caataatgac
      121 gtatgttccc atagtaacgc caatagggac tttccattga cgtcaatggg tggagtattt
      181 acggtaaact gcccacttgg cagtacatca agtgtatcat atgccaagta cgccccctat
      241 tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc cagtacatga ccttatggga
      301 ctttcctact tggcagtaca tctacgtatt agtcatcgct attaccatgg tgatgcggtt
      361 ttggcagtac atcaatgggc gtggatagcg gtttgactca cggggatttc caagtctcca
      421 ccccattgac gtcaatggga gtttgttttg gcaccaaaat caacgggact ttccaaaatg
      481 tcgtaacaac tccgccccat tgacgcaaat gggcggtagg cgtgtacggt gggaggtcta
      541 tataagcaga gctggtttag tgaaccgtca gatccgctag cgctaccgga ctcagatctc
      601 gagctcaagc ttcgaattct gcagtcgacg gtaccgcggg cccgggatcc accggtcgcc
      661 accatggtga gcaagggcga ggagctgttc accggggtgg tgcccatcct ggtcgagctg
      721 gacggcgacg taaacggcca caagttcagc gtgtccggcg agggcgaggg cgatgccacc
      781 tacggcaagc tgaccctgaa gttcatctgc accaccggca agctgcccgt gccctggccc
      841 accctcgtga ccaccctgac ctacggcgtg cagtgcttca gccgctaccc cgaccacatg
      901 aagcagcacg acttcttcaa gtccgccatg cccgaaggct acgtccagga gcgcaccatc
      961 ttcttcaagg acgacggcaa ctacaagacc cgcgccgagg tgaagttcga gggcgacacc
     1021 ctggtgaacc gcatcgagct gaagggcatc gacttcaagg aggacggcaa catcctgggg
     1081 cacaagctgg agtacaacta caacagccac aacgtctata tcatggccga caagcagaag
     1141 aacggcatca aggtgaactt caagatccgc cacaacatcg aggacggcag cgtgcagctc
     1201 gccgaccact accagcagaa cacccccatc ggcgacggcc ccgtgctgct gcccgacaac
     1261 cactacctga gcacccagtc cgccctgagc aaagacccca acgagaagcg cgatcacatg
     1321 gtcctgctgg agttcgtgac cgccgccggg atcactctcg gcatggacga gctgtacaag
     1381 taaagcggcc gcactcctca ggtgcatgcc tctcctccct ctctggcagg gccccatcct
     1441 gggcagaggg gcctggggct gggcccagag tccagccgtc cagctgctcc tttcccagtt
     1501 tgatttcaat aaatctggcc actccccttt tggtgggttg aacgttttaa cagccaaggg
     1561 tgcatccttc atggtctggg cttgcgtctg tcttggggga cttattcgtc ctggctctct
     1621 ttggtccttg ctctggtggg acatggaggc aagtgttgaa ggctgcctat cagaaggtgg
     1681 tggctggtgt ggccaatgcc ctggctcaca aataccactg agatcttttt ccctctgcca
     1741 aaaattatgg ggacatcatg aagccccttg agcatctgac ttctggctaa taaaggaaat
     1801 ttattttcat tgcaatagtg tgttggaatt ttttgtgtct ctcactcgga aggacatatg
     1861 ggagggcaaa tcatttaaaa catcagaatg agtatttggt ttagagtttg gcaacatatg
     1921 cccatatgct ggctgccatg aacaaaggtt ggctataaag aggtcatcag tatatgaaac
     1981 agccccctgc tgtccattcc ttattccata gaaaagcctt gacttgaggt tagatttttt
     2041 ttatattttg ttttgtgtta tttttttctt taacatccct aaaattttcc ttacatgttt
     2101 tactagccag atttttcctc ctctcctgac tactcccagt catagctgtc cctcttctct
     2161 tatggagatc cctcgacctg cagcccaagc ttggcgtaat catggtcata gctgtttcct
     2221 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag cataaagtgt
     2281 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     2341 gctttccagt cgggaaacct gtcgtgccag cggatccgca tctcaattag tcagcaacca
     2401 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     2461 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tcggcctctg
     2521 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctaa
     2581 cttgtttatt gcagcttata atggttacaa ataaagcaat agcatcacaa atttcacaaa
     2641 taaagcattt ttttcactgc attctagttg tggtttgtcc aaactcatca atgtatctta
     2701 tcatgtctgg atccgctgca ttaatgaatc ggccaacgcg cggggagagg cggtttgcgt
     2761 attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt tcggctgcgg
     2821 cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc aggggataac
     2881 gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa aaaggccgcg
     2941 ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa tcgacgctca
     3001 agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc ccctggaagc
     3061 tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc cgcctttctc
     3121 ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag ttcggtgtag
     3181 gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga ccgctgcgcc
     3241 ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc gccactggca
     3301 gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac agagttcttg
     3361 aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg cgctctgctg
     3421 aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca aaccaccgct
     3481 ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa aggatctcaa
     3541 gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa ctcacgttaa
     3601 gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt aaattaaaaa
     3661 tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag ttaccaatgc
     3721 ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat agttgcctga
     3781 ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc cagtgctgca
     3841 atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa ccagccagcc
     3901 ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca gtctattaat
     3961 tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa cgttgttgcc
     4021 attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt cagctccggt
     4081 tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc ggttagctcc
     4141 ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact catggttatg
     4201 gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc tgtgactggt
     4261 gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg ctcttgcccg
     4321 gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct catcattgga
     4381 aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc cagttcgatg
     4441 taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag cgtttctggg
     4501 tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac acggaaatgt
     4561 tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg ttattgtctc
     4621 atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt tccgcgcaca
     4681 tttccccgaa aagtgccacc tgggtcgaca ttgattattg actagtaata gta
//
