LOCUS       sequence-274594-        4733 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Randomly picked clone 4 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-4
FEATURES             Location/Qualifiers
     source          1..4733
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     273..292
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     532..553
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      595..604
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     misc_feature    606..686
                     /label=MCS
                     /note="multiple cloning site"
     promoter        complement(709..912)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(738..758)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        913..1288
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        1319..1423
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1424..1492,1493..2284)
                     /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(1642..1661)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2455..3043
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2944..2963
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3197..3214
                     /label=L4440
                     /note="L4440 vector, forward primer"
     polyA_signal    3282..3416
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3306..3325)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3360..3379
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(3422..3617)
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      3436..3571
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     protein_bind    3676..3697
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3712..3717,3718..3735,3736..3742)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3750..3766
                     /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     3755..3777
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3774..3790
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3774..3790
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4133..4152
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     polyA_signal    4151..4206
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     4252..4271
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     CDS             complement(join(4612..4733,1..592,593..595,596..598))
                     /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(4615..4733,1..592,593..595,596..598))
                     /codon_start=1
                     /product="enhanced GFP"
                     /label=EGFP
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(4657..4678)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
ORIGIN
        1 ggttgtcggg cagcagcacg gggccgtcgc cgatgggggt gttctgctgg tagtggtcgg
       61 cgagctgcac gctgccgtcc tcgatgttgt ggcggatctt gaagttcacc ttgatgccgt
      121 tcttctgctt gtcggccatg atatagacgt tgtggctgtt gtagttgtac tccagcttgt
      181 gccccaggat gttgccgtcc tccttgaagt cgatgccctt cagctcgatg cggttcacca
      241 gggtgtcgcc ctcgaacttc acctcggcgc gggtcttgta gttgccgtcg tccttgaaga
      301 agatggtgcg ctcctggacg tagccttcgg gcatggcgga cttgaagaag tcgtgctgct
      361 tcatgtggtc ggggtagcgg ctgaagcact gcacgccgta ggtcagggtg gtcacgaggg
      421 tgggccaggg cacgggcagc ttgccggtgg tgcagatgaa cttcagggtc agcttgccgt
      481 aggtggcatc gccctcgccc tcgccggaca cgctgaactt gtggccgttt acgtcgccgt
      541 ccagctcgac caggatgggc accaccccgg tgaacagctc ctcgcccttg ctcaccatgg
      601 tggcgaccgg tggatcccgg gcccgcggta ccgtcgactg cagaattcga agcttgagct
      661 cgagatctga gtccggtagc gctagcggat ctgacggttc actaaaccag ctctgcttat
      721 atagacctcc caccgtacac gcctaccgcc catttgcgtc aatggggcgg agttgttacg
      781 acattttgga aagtcccgtt gattttggtg ccaaaacaaa ctcccattga cgtcaatggg
      841 gtggagactt ggaaatcccc gtgagtcaaa ccgctatcca cgcccattga tgtactgcca
      901 aaaccgcatc accatggtaa tagcgatgac taatacgtag atgtactgcc aagtaggaaa
      961 gtcccataag gtcatgtact gggcataatg ccaggcgggc catttaccgt cattgacgtc
     1021 aatagggggc gtacttggca tatgatacac ttgatgtact gccaagtggg cagtttaccg
     1081 taaatactcc acccattgac gtcaatggaa agtccctatt ggcgttacta tgggaacata
     1141 cgtcattatt gacgtcaatg ggcgggggtc gttgggcggt cagccaggcg ggccatttac
     1201 cgtaagttat gtaacgcgga actccatata tgggctatga actaatgacc ccgtaattga
     1261 ttactattac tagtcaataa tcaatgtcga cccaggtggc acttttcggg gaaatgtgcg
     1321 cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc tcatgagaca
     1381 ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta ttcaacattt
     1441 ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg ctcacccaga
     1501 aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg gttacatcga
     1561 actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac gttttccaat
     1621 gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg acgccgggca
     1681 agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt actcaccagt
     1741 cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg ctgccataac
     1801 catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac cgaaggagct
     1861 aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt gggaaccgga
     1921 gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag caatggcaac
     1981 aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc aacaattaat
     2041 agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc ttccggctgg
     2101 ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta tcattgcagc
     2161 actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg ggagtcaggc
     2221 aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga ttaagcattg
     2281 gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac ttcattttta
     2341 atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa tcccttaacg
     2401 tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga
     2461 tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt
     2521 ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg gcttcagcag
     2581 agcgcagata ccaaatactg ttcttctagt gtagccgtag ttaggccacc acttcaagaa
     2641 ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg ctgctgccag
     2701 tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg ataaggcgca
     2761 gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac
     2821 cgaactgaga tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa
     2881 ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc
     2941 agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg
     3001 tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc
     3061 ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc
     3121 ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag
     3181 ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa
     3241 accgcctctc cccgcgcgtt ggccgattca ttaatgcagc ggatccagac atgataagat
     3301 acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc tttatttgtg
     3361 aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa caagttagct
     3421 ttttgcaaaa gcctaggcct ccaaaaaagc ctcctcacta cttctggaat agctcagagg
     3481 ccgaggcggc ctcggcctct gcataaataa aaaaaattag tcagccatgg ggcggagaat
     3541 gggcggaact gggcggagtt aggggcggga tgggcggagt taggggcggg actatggttg
     3601 ctgactaatt gagatgcgga tccgctggca cgacaggttt cccgactgga aagcgggcag
     3661 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     3721 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     3781 cagctatgac catgattacg ccaagcttgg gctgcaggtc gagggatctc cataagagaa
     3841 gagggacagc tatgactggg agtagtcagg agaggaggaa aaatctggct agtaaaacat
     3901 gtaaggaaaa ttttagggat gttaaagaaa aaaataacac aaaacaaaat ataaaaaaaa
     3961 tctaacctca agtcaaggct tttctatgga ataaggaatg gacagcaggg ggctgtttca
     4021 tatactgatg acctctttat agccaacctt tgttcatggc agccagcata tgggcatatg
     4081 ttgccaaact ctaaaccaaa tactcattct gatgttttaa atgatttgcc ctcccatatg
     4141 tccttccgag tgagagacac aaaaaattcc aacacactat tgcaatgaaa ataaatttcc
     4201 tttattagcc agaagtcaga tgctcaaggg gcttcatgat gtccccataa tttttggcag
     4261 agggaaaaag atctcagtgg tatttgtgag ccagggcatt ggccacacca gccaccacct
     4321 tctgataggc agccttcaac acttgcctcc atgtcccacc agagcaagga ccaaagagag
     4381 ccaggacgaa taagtccccc aagacagacg caagcccaga ccatgaagga tgcacccttg
     4441 gctgttaaaa cgttcacccc gacaaaaggg gagtgaacag atttattgaa atcaaactgg
     4501 gaaaggagca gctggacggc tggactctgg gcccagcccc aggcccctct gcccaggatg
     4561 gggccctgcc agagagggag gagaggcatg cacctgagga gtgcggccgc tttacttgta
     4621 cagctcgtcc atgccgagag tgatcccggc ggcggtcacg aactccagca ggaccatgtg
     4681 atcgcgcttc tcgttggggt ctttgctcag ggcggactgg gtgctcaggt agt
//
