LOCUS       sequence-194154-        4378 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Expresses 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 4378)
  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 4378)
  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: pN-iRS3GG
FEATURES             Location/Qualifiers
     source          1..4378
                     /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        1233..1398
                     /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     1321..1340
                     /label=pBAD Forward
                     /note="For vectors with E. coli araBAD promoter, forward 
                     primer"
     RBS             2533..2544
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     primer_bind     complement(2581..2609)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3204..3224
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     terminator      3327..3413
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     rep_origin      complement(3578..4166)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3658..3677)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(4254..4348)
                     /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 ccgcgcaagg 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 gtcgacaaac caattgtcca tattgcatca
     1141 gacattgccg tcactgcgtc ttttactggc tcttctcgct aaccaaaccg gtaaccccgc
     1201 ttattaaaag cattctgtaa caaagcggga ccaaagccat gacaaaaacg cgtaacaaaa
     1261 gtgtctataa tcacggcaga aaagtccaca ttgattattt gcacggcgtc acactttgct
     1321 atgccatagc atttttatcc ataagattag cggatcctac ctgacgcttt ttatcgcaac
     1381 tctctactgt ttctccatag aattcgagac gagaaaagca gaaaaaacgc cgctgaacgc
     1441 ggcgttttgg attgtttgct gtttctttat ttaaactgtt tgaggaaacg cagatcgttt
     1501 tcgaagaatg aacgcaggtc ggtgacgccg taacgcaaca tagtcagacg ctccatcccc
     1561 atcccgaagc cgaaaccaga gtaaacttcc gggtcgatgc caacgttacg caacacgttc
     1621 ggatgcacca tcccgcagcc cagcacttcc agccatttac cgtttttacc catgacgtcc
     1681 acttctgcag aaggttcggt aaacgggaag taggaaggac ggaagcgaat ctgcaaatct
     1741 tcctcaaaga agttacgcag gaagtcgtgc agcgtgcctt tcaggttggt aaagctgatg
     1801 ttggtatcaa caatcagacc ttccatctga tggaacatcg gcgtgtgagt ctggtcgtag
     1861 tcgttacgat aaacacggcc aggcgcgatg atacgaatcg gtggctgctg ggctttcatg
     1921 gtgcggatct gtacgccaga ggtctgggta cgcagcaggc gggtagtgtc aaaccagaaa
     1981 gtgtcgtggt cagcgcgcgc cgggtggtga ccaggaatgt tcagagcatc gaagttatga
     2041 taatcgtctt cgatttccgg cccggttgcc acggtaaagc caagctcacc gaagaaactt
     2101 tcgatacggt cgatggtacg ggtaaccgga tgcagaccgc cgttttcaat gcgacgacct
     2161 ggcagagaga catcaatcgt ttccgccgcc agacgcgcat tcagtgcagc gctttccagt
     2221 tccgctttac gcgcattcag cgcctgctga acctgctctt tcgcttcgtt gataaccgca
     2281 ccagctgccg gacgctcttc tggcggcagc tcacgcaggg tcgtcatctg aagggttaag
     2341 tgcccttttt tacccaaata ttcgacgcgc acattatcta acgcggcaac atctgacgcc
     2401 tggctaatgg ccgccttcgc actggcaacc agttctgcga gatgtgacat ctagtatttc
     2461 ccctctttca aagcgtaaac aacgtataac ggcgtatgcg tctctaccat tcacctcttg
     2521 gatttgggta ttaaagagga gaaaggtacc atgagtaaag gagaagaact tttcactgga
     2581 gttgtcccaa ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt
     2641 ggagagggtg aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact
     2701 ggaaaactac ctgttccatg gccaacactt gtcactactt tcggttatgg tgttcaatgc
     2761 tttgcgagat acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa
     2821 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtgct
     2881 gaagtcaagt ttgaaggtga tacccttgtt aatagaatcg agttaaaagg tattgatttt
     2941 aaagaagatg gaaacattct tggacacaaa ttggaataca actataactc acacaatgta
     3001 tacatcatgg cagacaaaca aaagaatgga atcaaagtta acttcaaaat tagacacaac
     3061 attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat
     3121 ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat
     3181 cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca
     3241 catggcatgg atgatctcta caaataaaag cttgatatcc aattcctgca gcccggggga
     3301 tcccatggta cgcgtgctag aggcatcaaa taaaacgaaa ggctcagtcg aaagactggg
     3361 cctttcgttt tatctgttgt ttgtcggtga acgctctcct gagtaggaca aatccgccgc
     3421 cctagaccta gggcgttcgg ctgcggcgag cggtatcagc tcactcaaag gcggtaatac
     3481 ggttatccac agaatcaggg gataacgcag gaaagaacat gtgagcaaaa ggccagcaaa
     3541 aggccaggaa ccgtaaaaag gccgcgttgc tggcgttttt ccataggctc cgcccccctg
     3601 acgagcatca caaaaatcga cgctcaagtc agaggtggcg aaacccgaca ggactataaa
     3661 gataccaggc gtttccccct ggaagctccc tcgtgcgctc tcctgttccg accctgccgc
     3721 ttaccggata cctgtccgcc tttctccctt cgggaagcgt ggcgctttct catagctcac
     3781 gctgtaggta tctcagttcg gtgtaggtcg ttcgctccaa gctgggctgt gtgcacgaac
     3841 cccccgttca gcccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg
     3901 taagacacga cttatcgcca ctggcagcag ccactggtaa caggattagc agagcgaggt
     3961 atgtaggcgg tgctacagag ttcttgaagt ggtggcctaa ctacggctac actagaagga
     4021 cagtatttgg tatctgcgct ctgctgaagc cagttacctt cggaaaaaga gttggtagct
     4081 cttgatccgg caaacaaacc accgctggta gcggtggttt ttttgtttgc aagcagcaga
     4141 ttacgcgcag aaaaaaagga tctcaagaag atcctttgat cttttctacg gggtctgacg
     4201 ctcagtggaa cgaaaactca cgttaaggga ttttggtcat gactagtgct tggattctca
     4261 ccaataaaaa acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag
     4321 gtcattactg gatctatcaa caggagtcca agcgagctct cgaaccccag agtcccgc
//
