LOCUS       sequence-498177-        5110 bp    DNA     circular SYN 26-JAN-2026
DEFINITION  In vitro transcription (SP6 promotor) or mammalian expression (CMV 
            promotor) of a recorder containing 3 box-b hairpins.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5110)
  TITLE     Cifuentes Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5110)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 26, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pCS2_EYFP_3xBoxB_Recorder_SV40
FEATURES             Location/Qualifiers
     source          1..5110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        35..53
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     35..52
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     CDS             94..120
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     CDS             join(121..123,124..126,127..840)
                     /codon_start=1
                     /product="enhanced YFP"
                     /label=EYFP
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTL
                     KFICTTGKLPVPWPTLVTTFGYGLQCFARYPDHMKQHDFFKSAMPEGYVQERTIFFKDD
                     GNYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYIMADKQKNGIK
                     VNFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSYQSALSKDPNEKRDHMVLL
                     EFVTAAGITLGMDELYK"
     primer_bind     complement(166..187)
                     /label=EGFP-N
                     /note="EGFP, reverse primer"
     primer_bind     complement(427..446)
                     /label=EXFP-R
                     /note="For distinguishing EGFP variants, reverse primer"
     primer_bind     774..795
                     /label=EGFP-C
                     /note="EGFP, forward primer"
     protein_bind    927..945
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    971..989
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     protein_bind    995..1013
                     /label=boxB
                     /bound_moiety="lambda-N peptide"
                     /note="lambda N peptide binding site consisting of an RNA 
                     hairpin within the N-utliization (nut) site of the lambda 
                     bacteriophage transcript (Tan and Frankel, 1995)"
     polyA_signal    1162..1296
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1186..1205)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1240..1259
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1409..1429)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1409..1427)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1448..1464)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1448..1464)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1461..1483)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1472..1488
                     /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(1496..1502,1503..1520,1521..1526))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1541..1562
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1679..1696)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1850..2438)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1930..1949)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2609..3400,3401..3469))
                     /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     3232..3251
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3470..3574)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3600..4055)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3737..3758)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3949..3968
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4126..5110
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
ORIGIN
        1 cgccattctg cctggggacg tcggagcaag cttgatttag gtgacactat agaatacaag
       61 ctacttgttc tttttgcagg atccgccacc atgtatccgt acgacgtacc ggattatgcg
      121 atggtgagca agggcgagga gctgttcacc ggggtggtgc ccatcctggt cgagctggac
      181 ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg gcgagggcga tgccacctac
      241 ggcaagctga ccctgaagtt catctgcacc accggcaagc tgcccgtgcc ctggcccacc
      301 ctcgtgacca ccttcggcta cggcctgcag tgcttcgccc gctaccccga ccacatgaag
      361 cagcacgact tcttcaagtc cgccatgccc gaaggctacg tccaggagcg caccatcttc
      421 ttcaaggacg acggcaacta caagacccgc gccgaggtga agttcgaggg cgacaccctg
      481 gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg acggcaacat cctggggcac
      541 aagctggagt acaactacaa cagccacaac gtctatatca tggccgacaa gcagaagaac
      601 ggcatcaagg tgaacttcaa gatccgccac aacatcgagg acggcagcgt gcagctcgcc
      661 gaccactacc agcagaacac ccccatcggc gacggccccg tgctgctgcc cgacaaccac
      721 tacctgagct accagtccgc cctgagcaaa gaccccaacg agaagcgcga tcacatggtc
      781 ctgctggagt tcgtgaccgc cgccgggatc actctcggca tggacgagct gtacaagtag
      841 ctcgagcctc tagaactata gtgagtcgta ttacacactc tttccctaca cgacgctctt
      901 ccgatctgcc aacgctattc tggctggggc cctgaagaag ggccccgcga ctaagtccaa
      961 ctactaaact gggccctgaa gaagggccca tatagggccc tgaagaaggg cccctatcca
     1021 acgcaattct ggcacgtcca atccaatcca atccaatcca atccaatcca attaaattag
     1081 attagattag attagattag attagattag acaggccttg atcggaagag cacacgtctg
     1141 aagtccagtc actacgtagt agatccagac atgataagat acattgatga gtttggacaa
     1201 accacaacta gaatgcagtg aaaaaaatgc tttatttgtg aaatttgtga tgctattgct
     1261 ttatttgtaa ccattataag ctgcaataaa caagttaaca acaacaattg cattcatttt
     1321 atgtttcagg ttcaggggga ggtgtgggag gttttttaat tcgcggccgc ggcgccaatg
     1381 cattgggccc ggtacccagc ttttgttccc tttagtgagg gttaattgcg cgcttggcgt
     1441 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     1501 tacgagccgg gagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     1561 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     1621 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     1681 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     1741 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     1801 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     1861 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     1921 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     1981 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     2041 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     2101 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     2161 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     2221 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     2281 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     2341 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     2401 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     2461 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     2521 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     2581 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     2641 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     2701 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     2761 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     2821 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     2881 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     2941 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     3001 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     3061 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     3121 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     3181 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     3241 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     3301 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     3361 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     3421 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     3481 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     3541 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccaccta
     3601 aattgtaagc gttaatattt tgttaaaatt cgcgttaaat ttttgttaaa tcagctcatt
     3661 ttttaaccaa taggccgaaa tcggcaaaat cccttataaa tcaaaagaat agaccgagat
     3721 agggttgagt gttgttccag tttggaacaa gagtccacta ttaaagaacg tggactccaa
     3781 cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca ctacgtgaac catcacccta
     3841 atcaagtttt ttggggtcga ggtgccgtaa agcactaaat cggaacccta aagggagccc
     3901 ccgatttaga gcttgacggg gaaagccggc gaacgtggcg agaaaggaag ggaagaaagc
     3961 gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc acgctgcgcg taaccaccac
     4021 acccgccgcg cttaatgcgc cgctacaggg cgcgtcccat tcgccattca ggctgcgcaa
     4081 ctgttgggaa gggcgatcgg tgcgggcctc ttcgctatta cgccagtcga ccgccaattc
     4141 aatatggcgt atatggactc atgccaattc aatatggtgg atctggacct gtgccaattc
     4201 aatatggcgt atatggactc gtgccaattc aatatggtgg atctggaccc cagccaattc
     4261 aatatggcgg acttggcacc atgccaattc aatatggcgg acctggcact gtgccaactg
     4321 gggaggggtc tacttggcac ggtgccaagt ttgaggaggg gtcttggccc tgtgccaagt
     4381 ccgccatatt gaattggcat ggtgccaata atggcggcca tattggctat atgccaggat
     4441 caatatatag gcaatatcca atatggccct atgccaatat ggctattggc caggttcaat
     4501 actatgtatt ggccctatgc catatagtat tccatatatg ggttttccta ttgacgtaga
     4561 tagcccctcc caatgggcgg tcccatatac catatatggg gcttcctaat accgcccata
     4621 gccactcccc cattgacgtc aatggtctct atatatggtc tttcctattg acgtcatatg
     4681 ggcggtccta ttgacgtata tggcgcctcc cccattgacg tcaattacgg taaatggccc
     4741 gcctggctca atgcccattg acgtcaatag gaccacccac cattgacgtc aatgggatgg
     4801 ctcattgccc attcatatcc gttctcacgc cccctattga cgtcaatgac ggtaaatggc
     4861 ccacttggca gtacatcaat atctattaat agtaacttgg caagtacatt actattggaa
     4921 gtacgccagg gtacattggc agtactccca ttgacgtcaa tggcggtaaa tggcccgcga
     4981 tggctgccaa gtacatcccc attgacgtca atggggaggg gcaatgacgc aaatgggcgt
     5041 tccattgacg taaatgggcg gtaggcgtgc ctaatgggag gtctatataa gcaatgctcg
     5101 tttagggaac
//
