LOCUS       sequence-63590-e        5227 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 5227)
  AUTHORS   Zeng M, Kuzirian MS, Harper L, Paradis S, Nakayama T, Lau NC
  TITLE     Organic small hairpin RNAs (OshR): A do-it-yourself platform for 
            transgene-based gene silencing.
  JOURNAL   Methods. 2013 May 23. pii: S1046-2023(13)00148-5. doi: 
            10.1016/j.ymeth.2013.05.007.
  PUBMED    23707624
REFERENCE   2  (bases 1 to 5227)
  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: "Methods. 
            2013 May 23. pii: S1046-2023(13)00148-5. doi: 
            10.1016/j.ymeth.2013.05.007."
COMMENT     Sequence Label: pGSH0
FEATURES             Location/Qualifiers
     source          1..5227
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      complement(30..458)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(140..161)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     352..371
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     585..607
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     599..616
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     600..616
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        649..667
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        694..1678
                     /label=CMV IE94 promoter
                     /note="enhancer/promoter region of simian cytomegalovirus 
                     major immediate early transcription unit IE94"
     5'UTR           1717..1759
                     /label=Xenopus globin 5'-UTR
                     /note="translational enhancer from the 5'-UTR of the major 
                     beta-globin gene of Xenopus laevis"
     primer_bind     complement(1830..1858)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2687..2707
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     polyA_signal    2779..2913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2803..2822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     2857..2876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     complement(3016..3036)
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        complement(3016..3034)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     primer_bind     complement(3078..3094)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3078..3094)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3091..3113)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3102..3118
                     /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(3126..3132,3133..3150,3151..3156))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3171..3192
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3309..3326)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3480..4068)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3560..3579)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4239..5030,5031..5099))
                     /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     4862..4881
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5100..5204)
                     /gene="bla"
                     /label=AmpR promoter
ORIGIN
        1 ctaaattgta agcgttaata ttttgttaaa attcgcgtta aatttttgtt aaatcagctc
       61 attttttaac caataggccg aaatcggcaa aatcccttat aaatcaaaag aatagaccga
      121 gatagggttg agtgttgttc cagtttggaa caagagtcca ctattaaaga acgtggactc
      181 caacgtcaaa gggcgaaaaa ccgtctatca gggcgatggc ccactacgtg aaccatcacc
      241 ctaatcaagt tttttggggt cgaggtgccg taaagcacta aatcggaacc ctaaagggag
      301 cccccgattt agagcttgac ggggaaagcc ggcgaacgtg gcgagaaagg aagggaagaa
      361 agcgaaagga gcgggcgcta gggcgctggc aagtgtagcg gtcacgctgc gcgtaaccac
      421 cacacccgcc gcgcttaatg cgccgctaca gggcgcgtcc cattcgccat tcaggctgcg
      481 caactgttgg gaagggcgat cggtgcgggc ctcttcgcta ttacgccagc tggcgaaagg
      541 gggatgtgct gcaaggcgat taagttgggt aacgccaggg ttttcccagt cacgacgttg
      601 taaaacgacg gccagtgagc gcgtagggat aacagggtaa tgcgcgcgta atacgactca
      661 ctatagggcg aattgggtac cgggcccccc ctcgaccata gccaattcaa tatggcgtat
      721 atggactcat gccaattcaa tatggtggat ctggacctgt gccaattcaa tatggcgtat
      781 atggactcgt gccaattcaa tatggtggat ctggacccca gccaattcaa tatggcggac
      841 ttggcaccat gccaattcaa tatggcggac ttggcactgt gccaactggg gaggggtcta
      901 cttggcacgg tgccaagttt gaggaggggt cttggccctg tgccaagtcc gccatattga
      961 attggcatgg tgccaataat ggcggccata ttggctatat gccaggatca atatataggc
     1021 aatatccaat atggccctat gccaatatgg ctattggcca ggttcaatac tatgtattgg
     1081 ccctatgcca tatagtattc catatatggg ttttcctatt gacgtagata gcccctccca
     1141 atgggcggtc ccatatacca tatatggggc ttcctaatac cgcccatagc cactccccca
     1201 ttgacgtcaa tggtctctat atatggtctt tcctattgac gtcatatggg cggtcctatt
     1261 gacgtatatg gcgcctcccc cattgacgtc aattacggta aatggcccgc ctggctcaat
     1321 gcccattgac gtcaatagga ccacccacca ttgacgtcaa tgggatggct cattgcccat
     1381 tcatatccgt tctcacgccc cctattgacg tcaatgacgg taaatggccc acttggcagt
     1441 acatcaatat ctattaatag taacttggca agtacattac tattggaagt acgccagggt
     1501 acattggcag tactcccatt gacgtcaatg gcggtaaatg gcccgcgatg gctgccaagt
     1561 acatccccat tgacgtcaat ggggaggggc aatgacgcaa atgggcgttc cattgacgta
     1621 aatgggcggt aggcgtgcct aatgggaggt ctatataagc aatgctcgtt tagggaaccg
     1681 ccattctgcc tggggacgtc ggagcaagct agcttgcttg ttctttttgc agaagctcag
     1741 aataaacgct caactttggc agatctgaat tcctgcagcc cgggggatcc acagccacca
     1801 tgagtaaagg agaagaactt ttcactggag ttgtcccaat tcttgttgaa ttagatggtg
     1861 atgttaatgg gtacaaattt tctgtcagtg gagagggtga aggtgatgca acatacggaa
     1921 aacttaccct taaatttatt tgcactactg gaaaactacc tgttccatgg ccaacacttg
     1981 tcactactct cacttatggt gttcaatgct tttcaagata tccagatcat atgaagcggc
     2041 acgacttctt caagagcgcc atgcctgagg gatacgtgca ggagaggacc atcttcttca
     2101 aggacgacgg gaactacaag acacgtgctg aagtcaagtt tgagggagac accctcgtca
     2161 acaggatcga gcttaaggta agaaaagttc acatctgagt aggtagaata aaaagctgca
     2221 tgccttcgtc gaccttcatg catcttcacg cgtagcttcc cttaatacaa gtgagatgat
     2281 ggcataccat ctttcgggac tgagttgatg tgaagagttt ttctgttttg ctgatcactt
     2341 gtatattatg tgactaatag ttaaagtgcc aaaataaaat gtgggaacat tgaatgtgta
     2401 tcctacttaa gggaatcgat ttcaaggagg acggaaacat cctcggccac aagttggaat
     2461 acaactacaa ctcccacaac gtatacatca tggccgacaa gcaaaagaac ggcatcaaag
     2521 ccaacttcaa gacccgccac aacatcgaag acggcggcgt gcaactcgct gatcattatc
     2581 aacaaaatac tccaattggc gatgaccctg tccttttacc agacaaccat tacctgtcca
     2641 cacaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     2701 ttgtaacggc tgctgggatt acacatggca tggatgaact atacaaaaat ctagaactat
     2761 agtgagtcgt attacgtaga tccagacatg ataagataca ttgatgagtt tggacaaacc
     2821 acaactagaa tgcagtgaaa aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta
     2881 tttgtaacca ttataagctg caataaacaa gttaacaaca acaattgcat tcattttatg
     2941 tttcaggttc agggggaggt gtgggaggtt ttttaattcg cggccgccac cgcggtggag
     3001 ctccagcttt tgttcccttt agtgagggtt aattgcgcgc attaccctgt tatccctacg
     3061 cgcttggcgt aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt
     3121 ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc
     3181 taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc
     3241 cagctgcatt aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct
     3301 tccgcttcct cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca
     3361 gctcactcaa aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac
     3421 atgtgagcaa aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt
     3481 ttccataggc tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg
     3541 cgaaacccga caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc
     3601 tctcctgttc cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc
     3661 gtggcgcttt ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc
     3721 aagctgggct gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac
     3781 tatcgtcttg agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt
     3841 aacaggatta gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct
     3901 aactacggct acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc
     3961 ttcggaaaaa gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt
     4021 ttttttgttt gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg
     4081 atcttttcta cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc
     4141 atgagattat caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa
     4201 tcaatctaaa gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag
     4261 gcacctatct cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg
     4321 tagataacta cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga
     4381 gacccacgct caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag
     4441 cgcagaagtg gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa
     4501 gctagagtaa gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc
     4561 atcgtggtgt cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca
     4621 aggcgagtta catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg
     4681 atcgttgtca gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat
     4741 aattctctta ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc
     4801 aagtcattct gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg
     4861 gataataccg cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg
     4921 gggcgaaaac tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt
     4981 gcacccaact gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca
     5041 ggaaggcaaa atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata
     5101 ctcttccttt ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac
     5161 atatttgaat gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa
     5221 gtgccac
//
