LOCUS       sequence-194130-        4851 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses default target RNA (gI intron) and pheS cassette (in place
            of an antisense RNA probe, for golden gate cloning) in E. coli 
            (kanR)..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4851)
  AUTHORS   Vazquez-Anderson J, Mihailovic MK, Baldridge KC, Reyes KG, Haning K,
            Cho SH, Amador P, Powell WB, Contreras LM
  TITLE     Optimization of a novel biophysical model using large scale in vivo 
            antisense hybridization data displays improved prediction 
            capabilities of structurally accessible RNA regions.
  JOURNAL   Nucleic Acids Res. 2017 May 19;45(9):5523-5538. doi: 
            10.1093/nar/gkx115.
  PUBMED    28334800
REFERENCE   2  (bases 1 to 4851)
  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"; doi: "10"; journalName: 
            "Nucleic Acids Res"; date: "2017-05-19- 19"; volume: "45"; issue: 
            "9"; pages: "5523-5538"
COMMENT     Sequence Label: p-OiRS3GG
FEATURES             Location/Qualifiers
     source          1..4851
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             complement(1..795)
                     /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(112..131)
                     /label=Neo-F
                     /note="Neomycin resistance gene, forward primer"
     primer_bind     722..741
                     /label=Neo-R
                     /note="Neomycin resistance gene, reverse primer"
     promoter        complement(1511..1794)
                     /gene="araBAD"
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the 
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     primer_bind     complement(1569..1588)
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     promoter        join(1799..1817,1818..1823,1824..1840,1841..1846,
                     1847..1872)
                     /label=PLtetO-1 promoter
                     /note="modified phage lambda PL promoter with tet operator 
                     sites (Lutz and Bujard, 1997)"
     protein_bind    1799..1817
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1824..1842
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             3006..3017
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(3054..3082)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3677..3697
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3800..3886
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(4051..4639)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4131..4150)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4727..4821)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
ORIGIN
        1 tcagaagaac tcgtcaagaa ggcgatagaa ggcgatgcgc tgcgaatcgg gagcggcgat
       61 accgtaaagc acgaggaagc ggtcagccca ttcgccgcca agctcttcag caatatcacg
      121 ggtagccaac gctatgtcct gatagcggtc cgccacaccc agccggccac agtcgatgaa
      181 tccagaaaag cggccatttt ccaccatgat attcggcaag caggcatcgc catgggtcac
      241 gacgagatcc tcgccgtcgg gcatgcgcgc cttgagcctg gcgaacagtt cggctggcgc
      301 gagcccctga tgctcttcgt ccagatcatc ctgatcgaca agaccggctt ccatccgagt
      361 acgtgctcgc tcgatgcgat gtttcgcttg gtggtcgaat gggcaggtag ccggatcaag
      421 cgtatgcagc cgccgcattg catcagccat gatggatact ttctcggcag gagcaaggtg
      481 agatgacagg agatcctgcc ccggcacttc gcccaatagc agccagtccc ttcccgcttc
      541 agtgacaacg tcgagcacag ctgcgcaagg aacgcccgtc gtggccagcc acgatagccg
      601 cgctgcctcg tcctgcagtt cattcagggc accggacagg tcggtcttga caaaaagaac
      661 cgggcgcccc tgcgctgaca gccggaacac ggcggcatca gagcagccga ttgtctgttg
      721 tgcccagtca tagccgaata gcctctccac ccaagcggcc ggagaacctg cgtgcaatcc
      781 atcttgttca atcatgcgaa acgatcctca tcctgtctct tgatcagatc ttgatcccct
      841 gcgccatcag atccttggcg gcaagaaagc catccagttt actttgcagg gcttcccaac
      901 cttaccagag ggcgccccag ctggcaattc cgacgtcaaa gggcccaagg cccagtcttt
      961 cgactgagcc tttcgtttta tttgatgcct ggcagttccc tactctcgca tggggagacc
     1021 ccacactacc atcggcgcta cggcgtttca cttctgagtt cggcatgggg tcaggtggga
     1081 ccaccgcgct actgccgcca ggcatctaga cgagtactcc aaaactaatc aatatacttt
     1141 cgcatacaaa ttagttccca gcggctccag tgttgcatca cttcatccgc tagctcccat
     1201 taaggagagg tccgactata tcttatgaga agaatacatc ttccccgacc gacatttagt
     1261 ctgtgaactg catccatatc aacagaagat ctgttgactt aggacttggc tgcgtggtta
     1321 ggaccatgtc cgtcagctta ttaccatacc ctttgcaagg ccatctcaaa gtttcccctg
     1381 agacttggta ctgaacggct gttgacccct ttcccgcaat ttgacggtct tgccttttaa
     1441 accgatgcaa tctattggtt taaagactag ctaccaggtg catgcctgat aacttttccc
     1501 tccgtcgact atggagaaac agtagagagt tgcgataaaa agcgtcaggt aggatccgct
     1561 aatcttatgg ataaaaatgc tatggcatag caaagtgtga cgccgtgcaa ataatcaatg
     1621 tggacttttc tgccgtgatt atagacactt ttgttacgcg tttttgtcat ggctttggtc
     1681 ccgctttgtt acagaatgct tttaataagc ggggttaccg gtttggttag cgagaagagc
     1741 cagtaaaaga cgcagtgacg gcaatgtctg atgcaatatg gacaattggt ttctcgagtc
     1801 cctatcagtg atagagattg acatccctat cagtgataga gatactgagc acatcagcag
     1861 gacgcactga ccgaattcga gacgagaaaa gcagaaaaaa cgccgctgaa cgcggcgttt
     1921 tggattgttt gctgtttctt tatttaaact gtttgaggaa acgcagatcg ttttcgaaga
     1981 atgaacgcag gtcggtgacg ccgtaacgca acatagtcag acgctccatc cccatcccga
     2041 agccgaaacc agagtaaact tccgggtcga tgccaacgtt acgcaacacg ttcggatgca
     2101 ccatcccgca gcccagcact tccagccatt taccgttttt acccatgacg tccacttctg
     2161 cagaaggttc ggtaaacggg aagtaggaag gacggaagcg aatctgcaaa tcttcctcaa
     2221 agaagttacg caggaagtcg tgcagcgtgc ctttcaggtt ggtaaagctg atgttggtat
     2281 caacaatcag accttccatc tgatggaaca tcggcgtgtg agtctggtcg tagtcgttac
     2341 gataaacacg gccaggcgcg atgatacgaa tcggtggctg ctgggctttc atggtgcgga
     2401 tctgtacgcc agaggtctgg gtacgcagca ggcgggtagt gtcaaaccag aaagtgtcgt
     2461 ggtcagcgcg cgccgggtgg tgaccaggaa tgttcagagc atcgaagtta tgataatcgt
     2521 cttcgatttc cggcccggtt gccacggtaa agccaagctc accgaagaaa ctttcgatac
     2581 ggtcgatggt acgggtaacc ggatgcagac cgccgttttc aatgcgacga cctggcagag
     2641 agacatcaat cgtttccgcc gccagacgcg cattcagtgc agcgctttcc agttccgctt
     2701 tacgcgcatt cagcgcctgc tgaacctgct ctttcgcttc gttgataacc gcaccagctg
     2761 ccggacgctc ttctggcggc agctcacgca gggtcgtcat ctgaagggtt aagtgccctt
     2821 ttttacccaa atattcgacg cgcacattat ctaacgcggc aacatctgac gcctggctaa
     2881 tggccgcctt cgcactggca accagttctg cgagatgtga catctagtat ttcccctctt
     2941 tcaaagcgta aacaacgtat aacggcgtat gcgtctctac cattcacctc ttggatttgg
     3001 gtattaaaga ggagaaaggt accatgagta aaggagaaga acttttcact ggagttgtcc
     3061 caattcttgt tgaattagat ggtgatgtta atgggcacaa attttctgtc agtggagagg
     3121 gtgaaggtga tgcaacatac ggaaaactta cccttaaatt tatttgcact actggaaaac
     3181 tacctgttcc atggccaaca cttgtcacta ctttcggtta tggtgttcaa tgctttgcga
     3241 gatacccaga tcatatgaaa cagcatgact ttttcaagag tgccatgccc gaaggttatg
     3301 tacaggaaag aactatattt ttcaaagatg acgggaacta caagacacgt gctgaagtca
     3361 agtttgaagg tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag
     3421 atggaaacat tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca
     3481 tggcagacaa acaaaagaat ggaatcaaag ttaacttcaa aattagacac aacattgaag
     3541 atggaagcgt tcaactagca gaccattatc aacaaaatac tccaattggc gatggccctg
     3601 tccttttacc agacaaccat tacctgtcca cacaatctgc cctttcgaaa gatcccaacg
     3661 aaaagagaga ccacatggtc cttcttgagt ttgtaacagc tgctgggatt acacatggca
     3721 tggatgatct ctacaaataa aagcttgata tccaattcct gcagcccggg ggatcccatg
     3781 gtacgcgtgc tagaggcatc aaataaaacg aaaggctcag tcgaaagact gggcctttcg
     3841 ttttatctgt tgtttgtcgg tgaacgctct cctgagtagg acaaatccgc cgccctagac
     3901 ctagggcgtt cggctgcggc gagcggtatc agctcactca aaggcggtaa tacggttatc
     3961 cacagaatca ggggataacg caggaaagaa catgtgagca aaaggccagc aaaaggccag
     4021 gaaccgtaaa aaggccgcgt tgctggcgtt tttccatagg ctccgccccc ctgacgagca
     4081 tcacaaaaat cgacgctcaa gtcagaggtg gcgaaacccg acaggactat aaagatacca
     4141 ggcgtttccc cctggaagct ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg
     4201 atacctgtcc gcctttctcc cttcgggaag cgtggcgctt tctcatagct cacgctgtag
     4261 gtatctcagt tcggtgtagg tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt
     4321 tcagcccgac cgctgcgcct tatccggtaa ctatcgtctt gagtccaacc cggtaagaca
     4381 cgacttatcg ccactggcag cagccactgg taacaggatt agcagagcga ggtatgtagg
     4441 cggtgctaca gagttcttga agtggtggcc taactacggc tacactagaa ggacagtatt
     4501 tggtatctgc gctctgctga agccagttac cttcggaaaa agagttggta gctcttgatc
     4561 cggcaaacaa accaccgctg gtagcggtgg tttttttgtt tgcaagcagc agattacgcg
     4621 cagaaaaaaa ggatctcaag aagatccttt gatcttttct acggggtctg acgctcagtg
     4681 gaacgaaaac tcacgttaag ggattttggt catgactagt gcttggattc tcaccaataa
     4741 aaaacgcccg gcggcaaccg agcgttctga acaaatccag atggagttct gaggtcatta
     4801 ctggatctat caacaggagt ccaagcgagc tctcgaaccc cagagtcccg c
//
