LOCUS       sequence-405027-        8611 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Single reporter with secondary structure from splicing library..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8611)
  AUTHORS   Liao SE, Sudarshan M, Regev O
  TITLE     Deciphering RNA splicing logic with interpretable machine learning.
  JOURNAL   Proc Natl Acad Sci U S A. 2023 Oct 10;120(41):e2221165120. doi: 
            10.1073/pnas.2221165120. Epub 2023 Oct 5.
  PUBMED    37796983
REFERENCE   2  (bases 1 to 8611)
  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"; journalName: "Proc Natl 
            Acad Sci U S A."; date: "2023-10-10"; volume: "120(41):e2221165120. 
            doi"; pages: " 10.1073/pnas.2221165120. Epub 2023 Oct 5"
COMMENT     Sequence Label: pRegev_S1
FEATURES             Location/Qualifiers
     source          1..8611
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(join(94..807,808..810,811..813))
                     /codon_start=1
                     /product="the original enhanced GFP (Yang et al., 1996)"
                     /label=EGFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     ANFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(139..160)
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     primer_bind     488..507
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     747..768
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     regulatory      810..819
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     promoter        complement(821..1999)
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          830..1768
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(853..873)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     promoter        2030..2307
                     /label=chicken beta-actin promoter
     primer_bind     3248..3270
                     /label=pCAGGS-5
                     /note="Chimeric intron in CAG promoter, forward primer"
     primer_bind     3297..3316
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        3297..3315
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(4681..4698)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4687..4911
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      4957..5385
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5044..5063)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     5254..5275
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5394..5414)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        5399..5728
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5579..5714
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     5641..5660
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     promoter        5776..5823
                     /label=EM7 promoter
                     /note="synthetic bacterial promoter "
     CDS             5842..6216
                     /codon_start=1
                     /gene="Sh ble from Streptoalloteichus hindustanus"
                     /product="antibiotic-binding protein"
                     /label=BleoR
                     /note="confers resistance to bleomycin, phleomycin, and 
                     Zeocin(TM)"
                     /translation="MAKLTSAVPVLTARDVAGAVEFWTDRLGFSRDFVEDDFAGVVRDD
                     VTLFISAVQDQVVPDNTLAWVWVRGLDELYAEWSEVVSTNFRDASGPAMTEIGEQPWGR
                     EFALRDPAGNCVHFVAEEQD"
     polyA_signal    6346..6467
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(6383..6402)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     6437..6456
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(6516..6532)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(6516..6532)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(6529..6551)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    6540..6556
                     /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(6564..6570,6571..6588,6589..6594))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6609..6630
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(6747..6764)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(6918..7506)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(6998..7017)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(7677..8468,8469..8537))
                     /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     8300..8319
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(join(8538..8611,1..31))
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
       61 tcgacggatc gggagatccc agtgtggtgg tacttacttg tacagctcgt ccatgccgag
      121 agtgatcccg gcggcggtca cgaactccag caggaccatg tgatcgcgct tctcgttggg
      181 gtctttgctc agggcggact gggtgctcag gtagtggttg tcgggcagca gcacggggcc
      241 gtcgccgatg ggggtgttct gctggtagtg gtcggcgagc tgcacgctgc cgtcctcgat
      301 gttgtggcgg atcttgaagt tcgccttgat gccgttcttc tgcttgtcgg ccatgatata
      361 gacgttgtgg ctgttgtagt tgtactccag cttgtgcccc aggatgttgc cgtcctcctt
      421 gaagtcgatg cccttcagct cgatgcggtt caccagggtg tcgccctcga acttcacctc
      481 ggcgcgggtc ttgtagttgc cgtcgtcctt gaagaagatg gtgcgctcct ggacgtagcc
      541 ttcgggcatg gcggacttga agaagtcgtg ctgcttcatg tggtcggggt agcggctgaa
      601 gcactgcacg ccgtaggtca gggtggtcac gagggtgggc cagggcacgg gcagcttgcc
      661 ggtggtgcag atgaacttca gggtcagctt gccgtaggtg gcatcgccct cgccctcgcc
      721 agacacgctg aacttgtggc cgtttacgtc gccgtccagc tcgaccagga tgggcaccac
      781 cccggtgaac agctcctcgc ccttgctcac catggtggcc tcacgacacc tgaaatggaa
      841 gaaaaaaact ttgaaccact gtctgaggct tgagaatgaa ccaagatcca aactcaaaaa
      901 gggcaaattc caaggagaat tacatcaagt gccaagctgg cctaacttca gtctccaccc
      961 actcagtgtg gggaaactcc atcgcataaa acccctcccc ccaacctaaa gacgacgtac
     1021 tccaaaagct cgagaactaa tcgaggtgcc tggacggcgc ccggtactcc gtggagtcac
     1081 atgaagcgac ggctgaggac ggaaaggccc ttttcctttg tgtgggtgac tcacccgccc
     1141 gctctcccga gcgccgcgtc ctccattttg agctccctgc agcagggccg ggaagcggcc
     1201 atctttccgc tcacgcaact ggtgccgacc gggccagcct tgccgcccag ggcggggcga
     1261 tacacggcgg cgcgaggcca ggcaccagag caggccggcc agcttgagac tacccccgtc
     1321 cgattctcgg tggccgcgct cgcaggcccc gcctcgccga acatgtgcgc tgggacgcac
     1381 gggccccgtc gccgcccgcg gccccaaaaa ccgaaatacc agtgtgcaga tcttggcccg
     1441 catttacaag actatcttgc cagaaaaaaa gcgtcgcagc aggtcatcaa aaattttaaa
     1501 tggctagaga cttatcgaaa gcagcgagac aggcgcgaag gtgccaccag attcgcacgc
     1561 ggcggcccca gcgcccaggc caggcctcaa ctcaagcacg aggcgaaggg gctccttaag
     1621 cgcaaggcct cgaactctcc cacccacttc caacccgaag ctcgggatca agaatcacgt
     1681 actgcagcca ggtggaagta attcaaggca cgcaagggcc ataacccgta aagaggccag
     1741 gcccgcggga accacacacg gcacttacct gtgttctggc ggcaaacccg ttgcgaaaaa
     1801 gaacgttcac ggcgactact gcacttatat acggttctcc cccaccctcg ggaaaaaggc
     1861 ggagccagta cacgacatca ctttcccagt ttaccccgcg ccaccttctc taggcaccgg
     1921 ttcaattgcc gacccctccc cccaacttct cggggactgt gggcgatgtg cgctctgccc
     1981 actgacgggc accggagcct acgtaggaat taattcgcca gcacagtggt cgaggtgagc
     2041 cccacgttct gcttcactct ccccatctcc cccccctccc cacccccaat tttgtattta
     2101 tttatttttt aattattttg tgcagcgatg ggggcggggg gggggggggg gcgcgcgcca
     2161 ggcggggcgg ggcggggcga ggggcggggc ggggcgaggc ggagaggtgc ggcggcagcc
     2221 aatcagagcg gcgcgctccg aaagtttcct tttatggcga ggcggcggcg gcggcggccc
     2281 tataaaaagc gaagcgcgcg gcgggcggga gtcgctgcgc gctgccttcg ccccgtgccc
     2341 cgctccgccg ccgcctcgcg ccgcccgccc cggctctgac tgaccgcgtt actcccacag
     2401 gtgagcgggc gggacggccc ttctcctccg ggctgtaatt agcgcttggt ttaatgacgg
     2461 cttgtttctt ttctgtggct gcgtgaaagc cttgaggggc tccgggaggg ccctttgtgc
     2521 ggggggagcg gctcgggggg tgcgtgcgtg tgtgtgtgcg tggggagcgc cgcgtgcggc
     2581 tccgcgctgc ccggcggctg tgagcgctgc gggcgcggcg cggggctttg tgcgctccgc
     2641 agtgtgcgcg aggggagcgc ggccgggggc ggtgccccgc ggtgcggggg gggctgcgag
     2701 gggaacaaag gctgcgtgcg gggtgtgtgc gtgggggggt gagcaggggg tgtgggcgcg
     2761 tcggtcgggc tgcaaccccc cctgcacccc cctccccgag ttgctgagca cggcccggct
     2821 tcgggtgcgg ggctccgtac ggggcgtggc gcggggctcg ccgtgccggg cggggggtgg
     2881 cggcaggtgg gggtgccggg cggggcgggg ccgcctcggg ccggggaggg ctcgggggag
     2941 gggcgcggcg gcccccggag cgccggcggc tgtcgaggcg cggcgagccg cagccattgc
     3001 cttttatggt aatcgtgcga gagggcgcag ggacttcctt tgtcccaaat ctgtgcggag
     3061 ccgaaatctg ggaggcgccg ccgcaccccc tctagcgggc gcggggcgaa gcggtgcggc
     3121 gccggcagga aggaaatggg cggggagggc cttcgtgcgt cgccgcgccg ccgtcccctt
     3181 ctccctctcc agcctcgggg ctgtccgcgg ggggacggct gccttcgggg gggacggggc
     3241 agggcggggt tcggcttctg gcgtgtgacc ggcggctcta gcgtttaaac ttaagctaat
     3301 acgactcact atagggagct tggtaccgca acctcaaaca gacaccatgg tgcacctgac
     3361 tcctgaggag aagtctgccg ttactgccct gtggggcaag gtgaacgtgg atgaagttgg
     3421 tggtgaggcc ctgggcaggt tggtatcaag gttacaagac aggtttaagg agaccaatag
     3481 aaactgggca tatggagaca gagaagactc ttgggtttct gataggcact gactctctct
     3541 gcctatgtct ttctctgcca tccaggttga gtcccgctta ccattgcatt taagaaagcg
     3601 gccatacgcc gctaagaccc tactcttcag aataccagca ggtctgacta tgggaccctt
     3661 gatgttttct ttccccttct tttctatggt taagttcatg tcataggaag gggagaagta
     3721 acagggtaca gtttagaatg ggaaacagac gaatgattgc atcagtgtgg aagtctcagg
     3781 atcgttttag tttcttttat ttgctgttca taacaattgt tttcttttgt ttaattcttg
     3841 ctttcttttt ttttcttctc cgcaattttt actattatac ttaatgcctt aacattgtgt
     3901 ataacaaaag gaaatatctc tgagatacat taagtaactt aaaaaaaaac tttacacagt
     3961 ctgcctagta cattactatt tggaatatat gtgtgcttat ttgcatattc ataatctccc
     4021 tactttattt tcttttattt ttaattgata cataatcatt atacatattt atgggttaaa
     4081 gtgtaatgtt ttaatatcga tacacatatt gaccaaatca gggtaatttt gcatttgtaa
     4141 ttttaaaaaa tgctttcttc ttttaatata cttttttgtt tatcttattt ctaatacttt
     4201 ccctaatctc tttctttcag ggcaataatg atacaatgta tcatgcctct ttgcaccatt
     4261 ctaaagaata acagtgataa tttctgggtt aaggcaatag caatatttct gcatataaat
     4321 atttctgcat ataaattgta actgatgtaa gaggtttcat attgctaata gcagctacaa
     4381 tccagctacc attctgcttt tattttatgg ttgggataag gctggattat tctgagtcca
     4441 agctaggccc ttttgctaat catgttcata cctcttatct tcctcccaca gctcctgggc
     4501 aacgtgctgg tctgtgtgct ggcccatcac tttggcaaag aattcacccc accagtgcag
     4561 gctgcctatc agaaagtggt ggctggtgtg gctaatgccc tggcccacaa gtatcactaa
     4621 gctcgctcta gactagacga ttaaattaca ttatagggcc cgtttaaacc cgctgatcag
     4681 cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct
     4741 tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc
     4801 attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg
     4861 aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg
     4921 cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa
     4981 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     5041 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     5101 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     5161 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     5221 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     5281 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     5341 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt
     5401 gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat
     5461 gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag
     5521 tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat
     5581 cccgccccta actccgccca gttccgccca ttctccgccc catggctgac taattttttt
     5641 tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg
     5701 cttttttgga ggcctaggct tttgcaaaaa gctcccggga gcttgtatat ccattttcgg
     5761 atctgatcag cacgtgttga caattaatca tcggcatagt atatcggcat agtataatac
     5821 gacaaggtga ggaactaaac catggccaag ttgaccagtg ccgttccggt gctcaccgcg
     5881 cgcgacgtcg ccggagcggt cgagttctgg accgaccggc tcgggttctc ccgggacttc
     5941 gtggaggacg acttcgccgg tgtggtccgg gacgacgtga ccctgttcat cagcgcggtc
     6001 caggaccagg tggtgccgga caacaccctg gcctgggtgt gggtgcgcgg cctggacgag
     6061 ctgtacgccg agtggtcgga ggtcgtgtcc acgaacttcc gggacgcctc cgggccggcc
     6121 atgaccgaga tcggcgagca gccgtggggg cgggagttcg ccctgcgcga cccggccggc
     6181 aactgcgtgc acttcgtggc cgaggagcag gactgacacg tgctacgaga tttcgattcc
     6241 accgccgcct tctatgaaag gttgggcttc ggaatcgttt tccgggacgc cggctggatg
     6301 atcctccagc gcggggatct catgctggag ttcttcgccc accccaactt gtttattgca
     6361 gcttataatg gttacaaata aagcaatagc atcacaaatt tcacaaataa agcatttttt
     6421 tcactgcatt ctagttgtgg tttgtccaaa ctcatcaatg tatcttatca tgtctgtata
     6481 ccgtcgacct ctagctagag cttggcgtaa tcatggtcat agctgtttcc tgtgtgaaat
     6541 tgttatccgc tcacaattcc acacaacata cgagccggaa gcataaagtg taaagcctgg
     6601 ggtgcctaat gagtgagcta actcacatta attgcgttgc gctcactgcc cgctttccag
     6661 tcgggaaacc tgtcgtgcca gctgcattaa tgaatcggcc aacgcgcggg gagaggcggt
     6721 ttgcgtattg ggcgctcttc cgcttcctcg ctcactgact cgctgcgctc ggtcgttcgg
     6781 ctgcggcgag cggtatcagc tcactcaaag gcggtaatac ggttatccac agaatcaggg
     6841 gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa aggccaggaa ccgtaaaaag
     6901 gccgcgttgc tggcgttttt ccataggctc cgcccccctg acgagcatca caaaaatcga
     6961 cgctcaagtc agaggtggcg aaacccgaca ggactataaa gataccaggc gtttccccct
     7021 ggaagctccc tcgtgcgctc tcctgttccg accctgccgc ttaccggata cctgtccgcc
     7081 tttctccctt cgggaagcgt ggcgctttct catagctcac gctgtaggta tctcagttcg
     7141 gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac cccccgttca gcccgaccgc
     7201 tgcgccttat ccggtaacta tcgtcttgag tccaacccgg taagacacga cttatcgcca
     7261 ctggcagcag ccactggtaa caggattagc agagcgaggt atgtaggcgg tgctacagag
     7321 ttcttgaagt ggtggcctaa ctacggctac actagaagaa cagtatttgg tatctgcgct
     7381 ctgctgaagc cagttacctt cggaaaaaga gttggtagct cttgatccgg caaacaaacc
     7441 accgctggta gcggtggttt ttttgtttgc aagcagcaga ttacgcgcag aaaaaaagga
     7501 tctcaagaag atcctttgat cttttctacg gggtctgacg ctcagtggaa cgaaaactca
     7561 cgttaaggga ttttggtcat gagattatca aaaaggatct tcacctagat ccttttaaat
     7621 taaaaatgaa gttttaaatc aatctaaagt atatatgagt aaacttggtc tgacagttac
     7681 caatgcttaa tcagtgaggc acctatctca gcgatctgtc tatttcgttc atccatagtt
     7741 gcctgactcc ccgtcgtgta gataactacg atacgggagg gcttaccatc tggccccagt
     7801 gctgcaatga taccgcgaga cccacgctca ccggctccag atttatcagc aataaaccag
     7861 ccagccggaa gggccgagcg cagaagtggt cctgcaactt tatccgcctc catccagtct
     7921 attaattgtt gccgggaagc tagagtaagt agttcgccag ttaatagttt gcgcaacgtt
     7981 gttgccattg ctacaggcat cgtggtgtca cgctcgtcgt ttggtatggc ttcattcagc
     8041 tccggttccc aacgatcaag gcgagttaca tgatccccca tgttgtgcaa aaaagcggtt
     8101 agctccttcg gtcctccgat cgttgtcaga agtaagttgg ccgcagtgtt atcactcatg
     8161 gttatggcag cactgcataa ttctcttact gtcatgccat ccgtaagatg cttttctgtg
     8221 actggtgagt actcaaccaa gtcattctga gaatagtgta tgcggcgacc gagttgctct
     8281 tgcccggcgt caatacggga taataccgcg ccacatagca gaactttaaa agtgctcatc
     8341 attggaaaac gttcttcggg gcgaaaactc tcaaggatct taccgctgtt gagatccagt
     8401 tcgatgtaac ccactcgtgc acccaactga tcttcagcat cttttacttt caccagcgtt
     8461 tctgggtgag caaaaacagg aaggcaaaat gccgcaaaaa agggaataag ggcgacacgg
     8521 aaatgttgaa tactcatact cttccttttt caatattatt gaagcattta tcagggttat
     8581 tgtctcatga gcggatacat atttgaatgt a
//
