LOCUS       sequence-482882-        8664 bp    DNA     circular SYN 15-AUG-2025
DEFINITION  Dual Color betaGlobin reporter to study splicing kinetics in vivo.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8664)
  AUTHORS   Coulon A, Ferguson ML, de Turris V, Palangat M, Chow CC, Larson DR
  TITLE     Kinetic competition during the transcription cycle results in 
            stochastic RNA processing.
  JOURNAL   Elife. 2014 Oct 1;3. doi: 10.7554/eLife.03939.
  PUBMED    25271374
REFERENCE   2  (bases 1 to 8664)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Elife."; 
            date: "2014-10-1"; volume: "3. doi"; pages: " 10.7554/eLife.03939"
COMMENT     Sequence Label: CFP_ betaGlobin_24XPP7_24XMS2_Splicing_ reporter
FEATURES             Location/Qualifiers
     source          1..8664
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             join(2870..2872,2873..2875,2876..3586)
                     /codon_start=1
                     /product="enhanced CFP"
                     /label=ECFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTWGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYISHNVYITADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(2915..2936)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(3176..3195)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     3523..3544
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     promoter        complement(4918..4936)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(4919..4936)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(4956..4973)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4962..5186
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      5232..5590
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5319..5338)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5529..5550
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5607..5626)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     5661..5680
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(5740..5756)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(5740..5756)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(5753..5775)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    5764..5780
                     /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(5788..5794,5795..5812,5813..5818))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5833..5854
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5971..5988)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6142..6730)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6222..6241)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(6901..7692,7693..7761))
                     /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     7524..7543
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(7762..7866)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7941..7960)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     protein_bind    join(8195..8213,8214..8236,8237..8255,8256..8278,8279..8297,
                     8298..8320,8321..8339,8340..8362,8363..8381,8382..8404,
                     8405..8423,8424..8446,8447..8465)
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        8498..8536
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     8533..8557
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
ORIGIN
        1 acatttgctt ctgacacaac tgtgttcact agcaacctca aacagacacc atggtgcacc
       61 tgactcctga ggagaagtct gccgttactg ccctgtgggg caaggtgaac gtggatgaag
      121 ttggtggtga ggccctgggc aggttggtat caaggttaca agacaggttt aaggagacca
      181 atagaaactg ggcatgtgga gacagagaag actcttgggt ttctgatagg cactgactct
      241 ctctgcctat tggtctattt tcccaccctt aggctgctgg tggtctaccc ttggacccag
      301 aggttctttg agtcctttgg ggatctgtcc actcctgatg ctgttatggg caaccctaag
      361 gtgaaggctc atggcaagaa agtgctcggt gcctttagtg atggcctggc tcacctggac
      421 aacctcaagg gcacctttgc cacactgagt gagctgcact gtgacaagct gcacgtggat
      481 cctgagaact tcagggtgag tctatgggac ccttgatgtt ttccttcccc ttcttttcta
      541 tggttaagtt catgtcatag gaaggggaga agtaacaggg tacagtttag aatgggaaac
      601 agacgaatga ttgcatcagt gtggaagtct caggatcgtt ttagtttctt ttatttgctg
      661 ttcataacaa ttgttttctt ttgtttaatt cttgctttct ttttttttct tctccgcaat
      721 ttttactatt atacttaatg ccttaacatt gtgtataaca aaaggaaata tctctgagat
      781 acattaagta acttaaaaaa aaactttaca cagtctgcct agtacattac tatttggaat
      841 atatgtgtgc ttatttgcat attcataatc tccctacttt attttctttt atttttaatt
      901 gatacataat cattatacat atttatgggt taaagtgtaa tgtatcgata ctcgagatcc
      961 taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct gcaggtcgac
     1021 tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt tcattagatc
     1081 ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc tgcaggtcga
     1141 ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg ttcattagat
     1201 cctaaggtac ctaattgcct agaaaggagc agacgatatg gcgtcgctcc ctgcaggtcg
     1261 actctagaaa ccagcagagc atatgggctc gctggctgca gtattcccgg gttcattaga
     1321 tcctaaggta cctaattgcc tagaaaggag cagacgatat ggcgtcgctc cctgcaggtc
     1381 gactctagaa accagcagag catatgggct cgctggctgc agtattcccg ggttcattag
     1441 atcctaaggt acctaattgc ctagaaagga gcagacgata tggcgtcgct ccctgcaggt
     1501 cgactctaga aaccagcaga gcatatgggc tcgctggctg cagtattccc gggttcatta
     1561 gatcctaagg tacctaattg cctagaaagg agcagacgat atggcgtcgc tccctgcagg
     1621 tcgactctag aaaccagcag agcatatggg ctcgctggct gcagtattcc cgggttcatt
     1681 agatcctaag gtacctaatt gcctagaaag gagcagacga tatggcgtcg ctccctgcag
     1741 gtcgactcta gaaaccagca gagcatatgg gctcgctggc tgcagtattc ccgggttcat
     1801 tagatcctaa ggtacctaat tgcctagaaa ggagcagacg atatggcgtc gctccctgca
     1861 ggtcgactct agaaaccagc agagcatatg ggctcgctgg ctgcagtatt cccgggttca
     1921 ttagatccta aggtacctaa ttgcctagaa aggagcagac gatatggcgt cgctccctgc
     1981 aggtcgactc tagaaaccag cagagcatat gggctcgctg gctgcagtat tcccgggttc
     2041 attagatcct aaggtaccta attgcctaga aaggagcaga cgatatggcg tcgctccctg
     2101 caggtcgact ctagaaacca gcagagcata tgggctcgct ggctgcagta ttcccgggtt
     2161 cattagatcc taaggtacct aattgcctag aaaggagcag acgatatggc gtcgctccct
     2221 gcaggtcgac tctagaaacc agcagagcat atgggctcgc tggctgcagt attcccgggt
     2281 tcattagatc ctaaggtacc taattgccta gaaaggagca gacgatatgg cgtcgctccc
     2341 tgcaggtcga ctctagaaac cagcagagca tatgggctcg ctggctgcag tattcccggg
     2401 ttcattagat ctcgagaacg cgtgtacaca tattgaccaa atcagggtaa ttttgcattt
     2461 gtaattttaa aaaatgcttt cttcttttaa tatacttttt tgtttatctt atttctaata
     2521 ctttccctaa tctctttctt tcagggcaat aatgatacaa tgtatcatgc ctctttgcac
     2581 cattctaaag aataacagtg ataatttctg ggttaaggca atagcaatat ttctgcatat
     2641 aaatatttct gcatataaat tgtaactgat gtaagaggtt tcatattgct aatagcagct
     2701 acaatccagc taccattctg cttttatttt atggttggga taaggctgga ttattctgag
     2761 tccaagctag gcccttttgc taatcatgtt catacctctt atcttcctcc cacagctcct
     2821 gggcaacgtg ctggtctgtg tgctggccca tcactttggc aaagaattca tggtgagcaa
     2881 gggcgaggag ctgttcaccg gggtggtgcc catcctggtc gagctggacg gcgacgtaaa
     2941 cggccacaag ttcagcgtgt ccggcgaggg cgagggcgat gccacctacg gcaagctgac
     3001 cctgaagttc atctgcacca ccggcaagct gcccgtgccc tggcccaccc tcgtgaccac
     3061 cctgacctgg ggcgtgcagt gcttcagccg ctaccccgac cacatgaagc agcacgactt
     3121 cttcaagtcc gccatgcccg aaggctacgt ccaggagcgc accatcttct tcaaggacga
     3181 cggcaactac aagacccgcg ccgaggtgaa gttcgagggc gacaccctgg tgaaccgcat
     3241 cgagctgaag ggcatcgact tcaaggagga cggcaacatc ctggggcaca agctggagta
     3301 caactacatc agccacaacg tctatatcac cgccgacaag cagaagaacg gcatcaaggc
     3361 caacttcaag atccgccaca acatcgagga cggcagcgtg cagctcgccg accactacca
     3421 gcagaacacc cccatcggcg acggccccgt gctgctgccc gacaaccact acctgagcac
     3481 ccagtccgcc ctgagcaaag accccaacga gaagcgcgat cacatggtcc tgctggagtt
     3541 cgtgaccgcc gccgggatca ctctcggcat ggacgagctg tacaagtcta agctatagat
     3601 ctgaattcta gaggatccta cggtacttat tgccaagaaa gcacgagcat cagccgtgcc
     3661 tccaggtcga atcttcaaac gacgacgatc acgcgtcgct ccagtattcc agggttcatc
     3721 agatcctacg gtacttattg ccaagaaagc acgagcatca gccgtgcctc caggtcgaat
     3781 cttcaaacga cgacgatcac gcgtcgctcc agtattccag ggttcatcag atcctacggt
     3841 acttattgcc aagaaagcac gagcatcagc cgtgcctcca ggtcgaatct tcaaacgacg
     3901 acgatcacgc gtcgctccag tattccaggg ttcatcagat cctacggtac ttattgccaa
     3961 gaaagcacga gcatcagccg tgcctccagg tcgaatcttc aaacgacgac gatcacgcgt
     4021 cgctccagta ttccagggtt catcagatcc tacggtactt attgccaaga aagcacgagc
     4081 atcagccgtg cctccaggtc gaatcttcaa acgacgacga tcacgcgtcg ctccagtatt
     4141 ccagggttca tcagatccta cggtacttat tgccaagaaa gcacgagcat cagccgtgcc
     4201 tccaggtcga atcttcaaac gacgacgatc acgcgtcgct ccagtattcc agggttcatc
     4261 agatcctacg gtacttattg ccaagaaagc acgagcatca gccgtgcctc caggtcgaat
     4321 cttcaaacga cgacgatcac gcgtcgctcc agtattccag ggttcatcag atcctacggt
     4381 acttattgcc aagaaagcac gagcatcagc cgtgcctcca ggtcgaatct tcaaacgacg
     4441 acgatcacgc gtcgctccag tattccaggg ttcatcagat cctacggtac ttattgccaa
     4501 gaaagcacga gcatcagccg tgcctccagg tcgaatcttc aaacgacgac gatcacgcgt
     4561 cgctccagta ttccagggtt catcagatcc tacggtactt attgccaaga aagcacgagc
     4621 atcagccgtg cctccaggtc gaatcttcaa acgacgacga tcacgcgtcg ctccagtatt
     4681 ccagggttca tcagatccta cggtacttat tgccaagaaa gcacgagcat cagccgtgcc
     4741 tccaggtcga atcttcaaac gacgacgatc acgcgtcgct ccagtattcc agggttcatc
     4801 agatcctacg gtacttattg ccaagaaagc acgagcatca gccgtgcctc caggtcgaat
     4861 cttcaaacga cgacgatcac gcgtcgctcc agtattccag ggttcatcag atcctattct
     4921 atagtgtcac ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca
     4981 gccatctgtt gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac
     5041 tgtcctttcc taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat
     5101 tctggggggt ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca
     5161 tgctggggat gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag
     5221 ggggtatccc cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     5281 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     5341 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     5401 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc
     5461 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     5521 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc
     5581 ttttgattta aatggttaca aataaagcaa tagcatcaca aatttcacaa ataaagcatt
     5641 tttttcactg cattctagtt gtggtttgtc caaactcatc aatgtatctt atcatgtctg
     5701 tataccgtcg acctctagct agagcttggc gtaatcatgg tcatagctgt ttcctgtgtg
     5761 aaattgttat ccgctcacaa ttccacacaa catacgagcc ggaagcataa agtgtaaagc
     5821 ctggggtgcc taatgagtga gctaactcac attaattgcg ttgcgctcac tgcccgcttt
     5881 ccagtcggga aacctgtcgt gccagctgca ttaatgaatc ggccaacgcg cggggagagg
     5941 cggtttgcgt attgggcgct cttccgcttc ctcgctcact gactcgctgc gctcggtcgt
     6001 tcggctgcgg cgagcggtat cagctcactc aaaggcggta atacggttat ccacagaatc
     6061 aggggataac gcaggaaaga acatgtgagc aaaaggccag caaaaggcca ggaaccgtaa
     6121 aaaggccgcg ttgctggcgt ttttccatag gctccgcccc cctgacgagc atcacaaaaa
     6181 tcgacgctca agtcagaggt ggcgaaaccc gacaggacta taaagatacc aggcgtttcc
     6241 ccctggaagc tccctcgtgc gctctcctgt tccgaccctg ccgcttaccg gatacctgtc
     6301 cgcctttctc ccttcgggaa gcgtggcgct ttctcatagc tcacgctgta ggtatctcag
     6361 ttcggtgtag gtcgttcgct ccaagctggg ctgtgtgcac gaaccccccg ttcagcccga
     6421 ccgctgcgcc ttatccggta actatcgtct tgagtccaac ccggtaagac acgacttatc
     6481 gccactggca gcagccactg gtaacaggat tagcagagcg aggtatgtag gcggtgctac
     6541 agagttcttg aagtggtggc ctaactacgg ctacactaga agaacagtat ttggtatctg
     6601 cgctctgctg aagccagtta ccttcggaaa aagagttggt agctcttgat ccggcaaaca
     6661 aaccaccgct ggtagcggtg gtttttttgt ttgcaagcag cagattacgc gcagaaaaaa
     6721 aggatctcaa gaagatcctt tgatcttttc tacggggtct gacgctcagt ggaacgaaaa
     6781 ctcacgttaa gggattttgg tcatgagatt atcaaaaagg atcttcacct agatcctttt
     6841 aaattaaaaa tgaagtttta aatcaatcta aagtatatat gagtaaactt ggtctgacag
     6901 ttaccaatgc ttaatcagtg aggcacctat ctcagcgatc tgtctatttc gttcatccat
     6961 agttgcctga ctccccgtcg tgtagataac tacgatacgg gagggcttac catctggccc
     7021 cagtgctgca atgataccgc gagacccacg ctcaccggct ccagatttat cagcaataaa
     7081 ccagccagcc ggaagggccg agcgcagaag tggtcctgca actttatccg cctccatcca
     7141 gtctattaat tgttgccggg aagctagagt aagtagttcg ccagttaata gtttgcgcaa
     7201 cgttgttgcc attgctacag gcatcgtggt gtcacgctcg tcgtttggta tggcttcatt
     7261 cagctccggt tcccaacgat caaggcgagt tacatgatcc cccatgttgt gcaaaaaagc
     7321 ggttagctcc ttcggtcctc cgatcgttgt cagaagtaag ttggccgcag tgttatcact
     7381 catggttatg gcagcactgc ataattctct tactgtcatg ccatccgtaa gatgcttttc
     7441 tgtgactggt gagtactcaa ccaagtcatt ctgagaatag tgtatgcggc gaccgagttg
     7501 ctcttgcccg gcgtcaatac gggataatac cgcgccacat agcagaactt taaaagtgct
     7561 catcattgga aaacgttctt cggggcgaaa actctcaagg atcttaccgc tgttgagatc
     7621 cagttcgatg taacccactc gtgcacccaa ctgatcttca gcatctttta ctttcaccag
     7681 cgtttctggg tgagcaaaaa caggaaggca aaatgccgca aaaaagggaa taagggcgac
     7741 acggaaatgt tgaatactca tactcttcct ttttcaatat tattgaagca tttatcaggg
     7801 ttattgtctc atgagcggat acatatttga atgtatttag aaaaataaac aaataggggt
     7861 tccgcgcaca tttccccgaa aagtgccacc tgacgtcgac ggatcgggag atctcccgat
     7921 cccctatggt gcactctcag tacaatctgc tctgatgccg catagttaag ccagtatctg
     7981 ctccctgctt gtgtgttgga ggtcgctgag tagtgcgcga gcaaaattta agctacaaca
     8041 aggcaaggct tgaccgacaa ttgcatgaag aatctgctta gggttaggcg ttttgcgctg
     8101 cttcgcgatg tacgggccag atatacgcgt tgacattgat tattgactag tagatctata
     8161 gcggccgctt gggtgcagat gtcgagttta ccactcccta tcagtgatag agaaaagtga
     8221 aagtcgagtt taccactccc tatcagtgat agagaaaagt gaaagtcgag tttaccactc
     8281 cctatcagtg atagagaaaa gtgaaagtcg agtttaccac tccctatcag tgatagagaa
     8341 aagtgaaagt cgagtttacc actccctatc agtgatagag aaaagtgaaa gtcgagttta
     8401 ccactcccta tcagtgatag agaaaagtga aagtcgagtt taccactccc tatcagtgat
     8461 agagaaaagt gaaagtcgag ctcggtaccc gggtcgaggt aggcgtgtac ggtgggaggc
     8521 ctatataagc agagctcgtt tagtgaaccg tcagatcgcc tggagacgcc atccacgctg
     8581 ttttgacctc catagaagac accgggaccg atccagcctc cgcggccccg aattcgagct
     8641 cggtacccgg ggataagctt gctt
//
