LOCUS       sequence-271779-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  EGFP followed by the poly(A) site from human CENPB gene.
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
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        4..379
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        380..583
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     534..554
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     misc_feature    606..686
                     /label=MCS
                     /note="multiple cloning site"
     regulatory      688..697
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             join(694..696,697..699,700..1413)
                     /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(694..696,697..699,700..1410)
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(739..760)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(1000..1019)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     1347..1368
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     complement(1754..1773)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    1819..1874
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(1873..1892)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(2235..2251)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(2235..2251)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(2248..2270)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    2259..2275
                     /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(2283..2289,2290..2307,2308..2313))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2328..2349
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        2408..2603
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2454..2589
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     polyA_signal    2609..2743
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2646..2665)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     2700..2719
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(2811..2828)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2982..3570)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3062..3081)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(3741..4532,4533..4601))
                     /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     4364..4383
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(4602..4706)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 gtcgacattg attattgact agtaatagta atcaattacg gggtcattag ttcatagccc
       61 atatatggag ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa
      121 cgacccccgc ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac
      181 tttccattga cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca
      241 agtgtatcat atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg
      301 gcattatgcc cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt
      361 agtcatcgct attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg
      421 gtttgactca cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg
      481 gcaccaaaat caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat
      541 gggcggtagg cgtgtacggt gggaggtcta tataagcaga gctggtttag tgaaccgtca
      601 gatccgctag cgctaccgga ctcagatctc gagctcaagc ttcgaattct gcagtcgacg
      661 gtaccgcggg cccgggatcc accggtcgcc accatggtga gcaagggcga ggagctgttc
      721 accggggtgg tgcccatcct ggtcgagctg gacggcgacg taaacggcca caagttcagc
      781 gtgtccggcg agggcgaggg cgatgccacc tacggcaagc tgaccctgaa gttcatctgc
      841 accaccggca agctgcccgt gccctggccc accctcgtga ccaccctgac ctacggcgtg
      901 cagtgcttca gccgctaccc cgaccacatg aagcagcacg acttcttcaa gtccgccatg
      961 cccgaaggct acgtccagga gcgcaccatc ttcttcaagg acgacggcaa ctacaagacc
     1021 cgcgccgagg tgaagttcga gggcgacacc ctggtgaacc gcatcgagct gaagggcatc
     1081 gacttcaagg aggacggcaa catcctgggg cacaagctgg agtacaacta caacagccac
     1141 aacgtctata tcatggccga caagcagaag aacggcatca aggtgaactt caagatccgc
     1201 cacaacatcg aggacggcag cgtgcagctc gccgaccact accagcagaa cacccccatc
     1261 ggcgacggcc ccgtgctgct gcccgacaac cactacctga gcacccagtc cgccctgagc
     1321 aaagacccca acgagaagcg cgatcacatg gtcctgctgg agttcgtgac cgccgccggg
     1381 atcactctcg gcatggacga gctgtacaag taaagcggcc gcactcctca ggtgcatgcc
     1441 tctcctccct ctctggcagg gccccatcct gggcagaggg gcctggggct gggcccagag
     1501 tccagccgtc cagctgctcc tttcccagtt tgatttcaat aaatctgtcc actccccttt
     1561 tgtgggggtg aacgttttaa cagccaaggg tgcatccttc atggtctggg cttgcgtctg
     1621 tcttggggga cttattcgtc ctggctctct ttggtccttg ctctggtggg acatggaggc
     1681 aagtgttgaa ggctgcctat cagaaggtgg tggctggtgt ggccaatgcc ctggctcaca
     1741 aataccactg agatcttttt ccctctgcca aaaattatgg ggacatcatg aagccccttg
     1801 agcatctgac ttctggctaa taaaggaaat ttattttcat tgcaatagtg tgttggaatt
     1861 ttttgtgtct ctcactcgga aggacatatg ggagggcaaa tcatttaaaa catcagaatg
     1921 agtatttggt ttagagtttg gcaacatatg cccatatgct ggctgccatg aacaaaggtt
     1981 ggctataaag aggtcatcag tatatgaaac agccccctgc tgtccattcc ttattccata
     2041 gaaaagcctt gacttgaggt tagatttttt ttatattttg ttttgtgtta tttttttctt
     2101 taacatccct aaaattttcc ttacatgttt tactagccag atttttcctc ctctcctgac
     2161 tactcccagt catagctgtc cctcttctct tatggagatc cctcgacctg cagcccaagc
     2221 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     2281 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     2341 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     2401 cggatccgca tctcaattag tcagcaacca tagtcccgcc cctaactccg cccatcccgc
     2461 ccctaactcc gcccagttcc gcccattctc cgccccatgg ctgactaatt ttttttattt
     2521 atgcagaggc cgaggccgcc tcggcctctg agctattcca gaagtagtga ggaggctttt
     2581 ttggaggcct aggcttttgc aaaaagctaa cttgtttatt gcagcttata atggttacaa
     2641 ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc attctagttg
     2701 tggtttgtcc aaactcatca atgtatctta tcatgtctgg atccgctgca ttaatgaatc
     2761 ggccaacgcg cggggagagg cggtttgcgt attgggcgct cttccgcttc ctcgctcact
     2821 gactcgctgc gctcggtcgt tcggctgcgg cgagcggtat cagctcactc aaaggcggta
     2881 atacggttat ccacagaatc aggggataac gcaggaaaga acatgtgagc aaaaggccag
     2941 caaaaggcca ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgcccc
     3001 cctgacgagc atcacaaaaa tcgacgctca agtcagaggt ggcgaaaccc gacaggacta
     3061 taaagatacc aggcgtttcc ccctggaagc tccctcgtgc gctctcctgt tccgaccctg
     3121 ccgcttaccg gatacctgtc cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc
     3181 tcacgctgta ggtatctcag ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac
     3241 gaaccccccg ttcagcccga ccgctgcgcc ttatccggta actatcgtct tgagtccaac
     3301 ccggtaagac acgacttatc gccactggca gcagccactg gtaacaggat tagcagagcg
     3361 aggtatgtag gcggtgctac agagttcttg aagtggtggc ctaactacgg ctacactaga
     3421 agaacagtat ttggtatctg cgctctgctg aagccagtta ccttcggaaa aagagttggt
     3481 agctcttgat ccggcaaaca aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag
     3541 cagattacgc gcagaaaaaa aggatctcaa gaagatcctt tgatcttttc tacggggtct
     3601 gacgctcagt ggaacgaaaa ctcacgttaa gggattttgg tcatgagatt atcaaaaagg
     3661 atcttcacct agatcctttt aaattaaaaa tgaagtttta aatcaatcta aagtatatat
     3721 gagtaaactt ggtctgacag ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc
     3781 tgtctatttc gttcatccat agttgcctga ctccccgtcg tgtagataac tacgatacgg
     3841 gagggcttac catctggccc cagtgctgca atgataccgc gagacccacg ctcaccggct
     3901 ccagatttat cagcaataaa ccagccagcc ggaagggccg agcgcagaag tggtcctgca
     3961 actttatccg cctccatcca gtctattaat tgttgccggg aagctagagt aagtagttcg
     4021 ccagttaata gtttgcgcaa cgttgttgcc attgctacag gcatcgtggt gtcacgctcg
     4081 tcgtttggta tggcttcatt cagctccggt tcccaacgat caaggcgagt tacatgatcc
     4141 cccatgttgt gcaaaaaagc ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag
     4201 ttggccgcag tgttatcact catggttatg gcagcactgc ataattctct tactgtcatg
     4261 ccatccgtaa gatgcttttc tgtgactggt gagtactcaa ccaagtcatt ctgagaatag
     4321 tgtatgcggc gaccgagttg ctcttgcccg gcgtcaatac gggataatac cgcgccacat
     4381 agcagaactt taaaagtgct catcattgga aaacgttctt cggggcgaaa actctcaagg
     4441 atcttaccgc tgttgagatc cagttcgatg taacccactc gtgcacccaa ctgatcttca
     4501 gcatctttta ctttcaccag cgtttctggg tgagcaaaaa caggaaggca aaatgccgca
     4561 aaaaagggaa taagggcgac acggaaatgt tgaatactca tactcttcct ttttcaatat
     4621 tattgaagca tttatcaggg ttattgtctc atgagcggat acatatttga atgtatttag
     4681 aaaaataaac aaataggggt tccgcgcaca tttccccgaa aagtgccacc tgg
//
