LOCUS       sequence-497428-        5027 bp    DNA     circular SYN 15-JAN-2026
DEFINITION  In vitro transcription (SP6 promotor) or mammalian expression (CMV 
            promotor) of an empty recorder containing an EcoRI site..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5027)
  TITLE     Cifuentes Lab Plasmids 2025
REFERENCE   2  (bases 1 to 5027)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Jan 15, 2026 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: pCS2_EYFP_EcoRI_SV40_Recorder
FEATURES             Location/Qualifiers
     source          1..5027
                     /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"
     polyA_signal    1079..1213
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1103..1122)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     1157..1176
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(1326..1346)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(1326..1344)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(1365..1381)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1365..1381)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1378..1400)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1389..1405
                     /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(1413..1419,1420..1437,1438..1443))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1458..1479
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(1596..1613)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(1767..2355)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(1847..1866)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2526..3317,3318..3386))
                     /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     3149..3168
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3387..3491)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(3517..3972)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3654..3675)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     3866..3885
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     promoter        4043..5027
                     /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 tggctgaatt ctatccaacg caattctggc acgtccaatc
      961 caatccaatc caatccaatc caatccaatt aaattagatt agattagatt agattagatt
     1021 agattagaca ggcctagatc ggaagagcac acgtctgaac tccagtcact acgtagtaga
     1081 tccagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     1141 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     1201 caataaacaa gttaacaaca acaattgcat tcattttatg tttcaggttc agggggaggt
     1261 gtgggaggtt ttttaattcg cggccgcggc gccaatgcat tgggcccggt acccagcttt
     1321 tgttcccttt agtgagggtt aattgcgcgc ttggcgtaat catggtcata gctgtttcct
     1381 gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccgggag cataaagtgt
     1441 aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg ctcactgccc
     1501 gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca acgcgcgggg
     1561 agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg
     1621 gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca
     1681 gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac
     1741 cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac
     1801 aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg
     1861 tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac
     1921 ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat
     1981 ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag
     2041 cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac
     2101 ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt
     2161 gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaagaac agtatttggt
     2221 atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc
     2281 aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga
     2341 aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac
     2401 gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt cacctagatc
     2461 cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta aacttggtct
     2521 gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct atttcgttca
     2581 tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg cttaccatct
     2641 ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga tttatcagca
     2701 ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt atccgcctcc
     2761 atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt taatagtttg
     2821 cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt tggtatggct
     2881 tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat gttgtgcaaa
     2941 aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc cgcagtgtta
     3001 tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc cgtaagatgc
     3061 ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat gcggcgaccg
     3121 agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag aactttaaaa
     3181 gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt accgctgttg
     3241 agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc ttttactttc
     3301 accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa gggaataagg
     3361 gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg aagcatttat
     3421 cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa taaacaaata
     3481 ggggttccgc gcacatttcc ccgaaaagtg ccacctaaat tgtaagcgtt aatattttgt
     3541 taaaattcgc gttaaatttt tgttaaatca gctcattttt taaccaatag gccgaaatcg
     3601 gcaaaatccc ttataaatca aaagaataga ccgagatagg gttgagtgtt gttccagttt
     3661 ggaacaagag tccactatta aagaacgtgg actccaacgt caaagggcga aaaaccgtct
     3721 atcagggcga tggcccacta cgtgaaccat caccctaatc aagttttttg gggtcgaggt
     3781 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
     3841 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
     3901 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
     3961 tacagggcgc gtcccattcg ccattcaggc tgcgcaactg ttgggaaggg cgatcggtgc
     4021 gggcctcttc gctattacgc cagtcgaccg ccaattcaat atggcgtata tggactcatg
     4081 ccaattcaat atggtggatc tggacctgtg ccaattcaat atggcgtata tggactcgtg
     4141 ccaattcaat atggtggatc tggaccccag ccaattcaat atggcggact tggcaccatg
     4201 ccaattcaat atggcggacc tggcactgtg ccaactgggg aggggtctac ttggcacggt
     4261 gccaagtttg aggaggggtc ttggccctgt gccaagtccg ccatattgaa ttggcatggt
     4321 gccaataatg gcggccatat tggctatatg ccaggatcaa tatataggca atatccaata
     4381 tggccctatg ccaatatggc tattggccag gttcaatact atgtattggc cctatgccat
     4441 atagtattcc atatatgggt tttcctattg acgtagatag cccctcccaa tgggcggtcc
     4501 catataccat atatggggct tcctaatacc gcccatagcc actcccccat tgacgtcaat
     4561 ggtctctata tatggtcttt cctattgacg tcatatgggc ggtcctattg acgtatatgg
     4621 cgcctccccc attgacgtca attacggtaa atggcccgcc tggctcaatg cccattgacg
     4681 tcaataggac cacccaccat tgacgtcaat gggatggctc attgcccatt catatccgtt
     4741 ctcacgcccc ctattgacgt caatgacggt aaatggccca cttggcagta catcaatatc
     4801 tattaatagt aacttggcaa gtacattact attggaagta cgccagggta cattggcagt
     4861 actcccattg acgtcaatgg cggtaaatgg cccgcgatgg ctgccaagta catccccatt
     4921 gacgtcaatg gggaggggca atgacgcaaa tgggcgttcc attgacgtaa atgggcggta
     4981 ggcgtgccta atgggaggtc tatataagca atgctcgttt agggaac
//
