LOCUS       sequence-199856-        5602 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5602)
  AUTHORS   Iioka H, Loiselle D, Haystead TA, Macara IG
  TITLE     Efficient detection of RNA-protein interactions using tethered RNAs.
  JOURNAL   Nucleic Acids Res. 2011 Apr;39(8):e53. Epub 2011 Feb 7.
  PUBMED    21300640
REFERENCE   2  (bases 1 to 5602)
  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: "Nucleic 
            Acids Res."; date: "2011-04"; volume: "39(8)"; pages: "e53. Epub 
            2011 Feb 7"
COMMENT     Sequence Label: pcDNA3-tRSA-G quartet
FEATURES             Location/Qualifiers
     source          1..5602
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        165..544
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        545..748
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     699..719
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     793..812
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        793..811
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(1085..1103)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1086..1103)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1123..1140)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1129..1353
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      1399..1827
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(1486..1505)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     1696..1717
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1836..1856)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1841..2170
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2021..2156
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2083..2102
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     CDS             2237..3031
                     /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(2291..2310)
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     primer_bind     2901..2920
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     polyA_signal    3205..3326
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3242..3261)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3296..3315
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3375..3391)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3375..3391)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3388..3410)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3399..3415
                     /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(3423..3429,3430..3447,3448..3453))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3468..3489
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3606..3623)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3777..4365)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3857..3876)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4536..5327,5328..5396))
                     /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     5159..5178
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5397..5501)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5576..5595)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 aagccagtat ctgctccctg cttgtgtgtt ggaggtcgct gagtagtgcg cgagcaaaat
       61 ttaagctaca acaaggcaag gcttgaccga caattgcatg aagaatctgc ttagggttag
      121 gcgttttgcg ctgcttcgcg atgtacgggc cagatatacg cgttgacatt gattattgac
      181 tagttattaa tagtaatcaa ttacggggtc attagttcat agcccatata tggagttccg
      241 cgttacataa cttacggtaa atggcccgcc tggctgaccg cccaacgacc cccgcccatt
      301 gacgtcaata atgacgtatg ttcccatagt aacgccaata gggactttcc attgacgtca
      361 atgggtggag tatttacggt aaactgccca cttggcagta catcaagtgt atcatatgcc
      421 aagtacgccc cctattgacg tcaatgacgg taaatggccc gcctggcatt atgcccagta
      481 catgacctta tgggactttc ctacttggca gtacatctac gtattagtca tcgctattac
      541 catggtgatg cggttttggc agtacatcaa tgggcgtgga tagcggtttg actcacgggg
      601 atttccaagt ctccacccca ttgacgtcaa tgggagtttg ttttggcacc aaaatcaacg
      661 ggactttcca aaatgtcgta acaactccgc cccattgacg caaatgggcg gtaggcgtgt
      721 acggtgggag gtctatataa gcagagctct ctggctaact agagaaccca ctgcttactg
      781 gcttatcgaa attaatacga ctcactatag ggagacccaa gcttggtacc gagctcggat
      841 ccactagtaa cggccgccag tgtgctggaa ttgaaaaaaa aaaaagcccg gatagctcag
      901 tcggtagagc agcggcctcg accagaatca tgcaagtgcg taagatagtc gcgggtcgag
      961 gccgcgtcca gggttcaagt ccctgttcgg gcgccactgc agaaaaaaaa aaaagaattc
     1021 ggctgcggtg tggaaggagt ggctgggttg cgcagctctc gagcatgcat ctagagggcc
     1081 ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact gtgccttcta
     1141 gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg gaaggtgcca
     1201 ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg agtaggtgtc
     1261 attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg gaagacaata
     1321 gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga accagctggg
     1381 gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg
     1441 ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct
     1501 tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc
     1561 ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt gattagggtg
     1621 atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt
     1681 ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg
     1741 tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc
     1801 tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt tagggtgtgg
     1861 aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc
     1921 aaccaggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa gcatgcatct
     1981 caattagtca gcaaccatag tcccgcccct aactccgccc atcccgcccc taactccgcc
     2041 cagttccgcc cattctccgc cccatggctg actaattttt tttatttatg cagaggccga
     2101 ggccgcctct gcctctgagc tattccagaa gtagtgagga ggcttttttg gaggcctagg
     2161 cttttgcaaa aagctcccgg gagcttgtat atccattttc ggatctgatc aagagacagg
     2221 atgaggatcg tttcgcatga ttgaacaaga tggattgcac gcaggttctc cggccgcttg
     2281 ggtggagagg ctattcggct atgactgggc acaacagaca atcggctgct ctgatgccgc
     2341 cgtgttccgg ctgtcagcgc aggggcgccc ggttcttttt gtcaagaccg acctgtccgg
     2401 tgccctgaat gaactgcagg acgaggcagc gcggctatcg tggctggcca cgacgggcgt
     2461 tccttgcgca gctgtgctcg acgttgtcac tgaagcggga agggactggc tgctattggg
     2521 cgaagtgccg gggcaggatc tcctgtcatc tcaccttgct cctgccgaga aagtatccat
     2581 catggctgat gcaatgcggc ggctgcatac gcttgatccg gctacctgcc cattcgacca
     2641 ccaagcgaaa catcgcatcg agcgagcacg tactcggatg gaagccggtc ttgtcgatca
     2701 ggatgatctg gacgaagagc atcaggggct cgcgccagcc gaactgttcg ccaggctcaa
     2761 ggcgcgcatg cccgacggcg aggatctcgt cgtgacccat ggcgatgcct gcttgccgaa
     2821 tatcatggtg gaaaatggcc gcttttctgg attcatcgac tgtggccggc tgggtgtggc
     2881 ggaccgctat caggacatag cgttggctac ccgtgatatt gctgaagagc ttggcggcga
     2941 atgggctgac cgcttcctcg tgctttacgg tatcgccgct cccgattcgc agcgcatcgc
     3001 cttctatcgc cttcttgacg agttcttctg agcgggactc tggggttcga aatgaccgac
     3061 caagcgacgc ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg
     3121 ttgggcttcg gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc
     3181 atgctggagt tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa
     3241 agcaatagca tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt
     3301 ttgtccaaac tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc
     3361 ttggcgtaat catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca
     3421 cacaacatac gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa
     3481 ctcacattaa ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag
     3541 ctgcattaat gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc
     3601 gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct
     3661 cactcaaagg cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg
     3721 tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc
     3781 cataggctcc gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga
     3841 aacccgacag gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct
     3901 cctgttccga ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg
     3961 gcgctttctc atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag
     4021 ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat
     4081 cgtcttgagt ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac
     4141 aggattagca gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac
     4201 tacggctaca ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc
     4261 ggaaaaagag ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt
     4321 tttgtttgca agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc
     4381 ttttctacgg ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg
     4441 agattatcaa aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca
     4501 atctaaagta tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca
     4561 cctatctcag cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag
     4621 ataactacga tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac
     4681 ccacgctcac cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc
     4741 agaagtggtc ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct
     4801 agagtaagta gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc
     4861 gtggtgtcac gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg
     4921 cgagttacat gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc
     4981 gttgtcagaa gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat
     5041 tctcttactg tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag
     5101 tcattctgag aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat
     5161 aataccgcgc cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg
     5221 cgaaaactct caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca
     5281 cccaactgat cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga
     5341 aggcaaaatg ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc
     5401 ttcctttttc aatattattg aagcatttat cagggttatt gtctcatgag cggatacata
     5461 tttgaatgta tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg
     5521 ccacctgacg tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa
     5581 tctgctctga tgccgcatag tt
//
