LOCUS       sequence-233984-        5749 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  ssRNA expression for dsRNA production.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5749)
  TITLE     Petr Svoboda lab unpublished
REFERENCE   2  (bases 1 to 5749)
  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"
COMMENT     Sequence Label: CMV-Lin28a-as
FEATURES             Location/Qualifiers
     source          1..5749
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        107..486
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        487..690
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     641..661
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     735..754
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        735..753
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1212..1229)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1218..1442
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1488..1916
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1575..1594)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1785..1806
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1925..1945)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1930..2259
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2110..2245
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2172..2191
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2326..3120
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /label=NeoR/KanR
                     /note="confers resistance to neomycin, kanamycin, and G418 
                     (Geneticin(R))"
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF"
     primer_bind     complement(2380..2399)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2990..3009
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3294..3415
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3331..3350)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3385..3404
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3464..3480)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3464..3480)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3477..3499)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3488..3504
                     /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(3512..3518,3519..3536,3537..3542))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3557..3578
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3695..3712)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3866..4454)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3946..3965)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4625..5416,5417..5485))
                     /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     5248..5267
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5486..5590)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5665..5684)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagggtt
       61 aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgttgaca ttgattattg
      121 actagttatt aatagtaatc aattacgggg tcattagttc atagcccata tatggagttc
      181 cgcgttacat aacttacggt aaatggcccg cctggctgac cgcccaacga cccccgccca
      241 ttgacgtcaa taatgacgta tgttcccata gtaacgccaa tagggacttt ccattgacgt
      301 caatgggtgg agtatttacg gtaaactgcc cacttggcag tacatcaagt gtatcatatg
      361 ccaagtacgc cccctattga cgtcaatgac ggtaaatggc ccgcctggca ttatgcccag
      421 tacatgacct tatgggactt tcctacttgg cagtacatct acgtattagt catcgctatt
      481 accatggtga tgcggttttg gcagtacatc aatgggcgtg gatagcggtt tgactcacgg
      541 ggatttccaa gtctccaccc cattgacgtc aatgggagtt tgttttggca ccaaaatcaa
      601 cgggactttc caaaatgtcg taacaactcc gccccattga cgcaaatggg cggtaggcgt
      661 gtacggtggg aggtctatat aagcagagct ctctggctaa ctagagaacc cactgcttac
      721 tggcttatcg aaattaatac gactcactat agggagaccc aagctggcta gcgtttaaac
      781 gggccctcta gactcgagcg gccgcggagt ttaaggaaag aggcatcaat ctgcagagtg
      841 gagaaagtgg gggtaagggt gggttgcgtg ggtagcttgc actgccgtgt ctcaggccgg
      901 ggttcccagt gtcaccctgt ctttccttgg agggaaggaa aggatgagac aaaggaactc
      961 ctaccacact ctatctgaaa gcaagtgaag gcttttgtgg ggaggaacca ccctagaacc
     1021 cgaggctttg ccaagtggct gggctaggga agttcttttg tagaaggctg tgtgatattt
     1081 cccttgccag acgggaagcg aaacaagtgt caaaccgcgg ccgccactgt gctggatatc
     1141 tgcagaattc caccacactg gactagtgga tccgagctcg gtaccaagct taagtttaaa
     1201 ccgctgatca gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc
     1261 cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga
     1321 aattgcatcg cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga
     1381 cagcaagggg gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat
     1441 ggcttctgag gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag
     1501 cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag
     1561 cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt
     1621 tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca
     1681 cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata
     1741 gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca
     1801 aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc
     1861 gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt
     1921 ctgtggaatg tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt
     1981 atgcaaagca tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca
     2041 gcaggcagaa gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta
     2101 actccgccca tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga
     2161 ctaatttttt ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag
     2221 tagtgaggag gcttttttgg aggcctaggc ttttgcaaaa agctcccggg agcttgtata
     2281 tccattttcg gatctgatca agagacagga tgaggatcgt ttcgcatgat tgaacaagat
     2341 ggattgcacg caggttctcc ggccgcttgg gtggagaggc tattcggcta tgactgggca
     2401 caacagacaa tcggctgctc tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg
     2461 gttctttttg tcaagaccga cctgtccggt gccctgaatg aactgcagga cgaggcagcg
     2521 cggctatcgt ggctggccac gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact
     2581 gaagcgggaa gggactggct gctattgggc gaagtgccgg ggcaggatct cctgtcatct
     2641 caccttgctc ctgccgagaa agtatccatc atggctgatg caatgcggcg gctgcatacg
     2701 cttgatccgg ctacctgccc attcgaccac caagcgaaac atcgcatcga gcgagcacgt
     2761 actcggatgg aagccggtct tgtcgatcag gatgatctgg acgaagagca tcaggggctc
     2821 gcgccagccg aactgttcgc caggctcaag gcgcgcatgc ccgacggcga ggatctcgtc
     2881 gtgacccatg gcgatgcctg cttgccgaat atcatggtgg aaaatggccg cttttctgga
     2941 ttcatcgact gtggccggct gggtgtggcg gaccgctatc aggacatagc gttggctacc
     3001 cgtgatattg ctgaagagct tggcggcgaa tgggctgacc gcttcctcgt gctttacggt
     3061 atcgccgctc ccgattcgca gcgcatcgcc ttctatcgcc ttcttgacga gttcttctga
     3121 gcgggactct ggggttcgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     3181 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     3241 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     3301 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3361 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3421 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3481 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3541 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3601 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3661 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3721 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3781 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3841 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3901 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3961 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     4021 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     4081 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     4141 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     4201 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     4261 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     4321 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4381 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4441 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4501 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4561 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4621 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4681 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4741 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4801 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4861 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4921 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4981 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     5041 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     5101 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     5161 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     5221 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     5281 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     5341 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5401 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5461 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5521 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5581 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5641 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5701 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaa
//
