LOCUS       sequence-12963-e        8059 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 8059)
  AUTHORS   Zhang H, Fire AZ
  TITLE     Cell autonomous specification of temporal identity by Caenorhabditis
            elegans microRNA lin-4.
  JOURNAL   Dev Biol. 2010 Aug 15. 344(2):603-10.
  PUBMED    20493184
REFERENCE   2  (bases 1 to 8059)
  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: "Dev Biol. 
            2010 Aug 15. 344(2):603-10."
COMMENT     Sequence Label: pHZ054
FEATURES             Location/Qualifiers
     source          1..8059
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1988..2010)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2110..2129
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2207..2225)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2293..2397
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2398..2466,2467..3258)
                     /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     complement(2616..2635)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3429..4017
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3918..3937
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4171..4188
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4305..4326
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4341..4346,4347..4364,4365..4371)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4379..4395
                     /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)."
     primer_bind     4384..4406
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4403..4419
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4403..4419
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             6848..6868
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6869..6889
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6890..6910
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6911..6931
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             6932..6952
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
ORIGIN
        1 aagtggaatc acaatctcct cctcttcaag gtccacaata gaatgccaat ttttcgagtc
       61 atccttcggg caatgttcat tacactttct ctctgttgta cttgagcatg tttcaatttc
      121 aatcacaaat gccttttttg gagagaattg aaggcaaaac caaactaaat tgagtttggt
      181 ggaaattgaa attgcaacat gctcaattca ttttgttttt ttcttttaac tatatggatg
      241 ccacgctgga ttgactcttc cgtacactca cgctcattcc aaattatccc catctgaccc
      301 cggactgttt tatccaaacc tatactgatg tatattgtat gcggcatgtt attttttcat
      361 aaccacaagc atttagcgct ttcggcttca gatttctaat cgctgtttct ttactgcttt
      421 gccctttttt ctaacttctg aatttcgtta ttatgcaaca attctacctc aatttttttg
      481 tatctttccc caattctagt tgacattatt ggttttgatt gttttcctag ctctgaaatc
      541 ccacaactag gcgccactga attgtatctt tcattcccac tttcgttttt cgcacattgt
      601 agtatctctc aattaggaaa cacttgaaaa cctctattgc ttaggattga tgagagatat
      661 tgctttttcc tgcactcact ttacctttgt ctcacctcaa aaattgctct caggaacatt
      721 caaaactcag gaatttgtca ccttggtctc tcatcatata tcttacctct tgtcacaccc
      781 cccatcccca gtctcaagtt catttcttac tttgtaactc cgtttagtgc gcccaattct
      841 cgtcattttg attacactct cttttaatcc aactcaggga accaattttt tttctcattg
      901 aactcaggaa tttcttctac ctcagggaac ctacctcatc cacttttcag ttgtttgggg
      961 ccaaatatct atatccaaag tagtagtcta caatttagta ttttattatt acctcccgcc
     1021 gttttagctt ttaatgttaa aatcaggaac ttttgaaaat gatcttcacc tcattcagaa
     1081 gcaaaaatca ggcattttcc aaagattttg aaaacacata aacctccttc caagtcaaaa
     1141 ctcacaacca actcagggac ctttttctta cttctgtatc acaaaaatga ttatatttct
     1201 gatgaatgtt gctctgtcat aaatcaattt atttcttttg aaccgaaagc cgaaatgtgt
     1261 ttttcctatt tgtcgtcatt gttctcatcc accccaactt cctctcatta ttgaagtttc
     1321 tcatgtattc tacctcattt attattgttt ttttttgttc aaaatgtcaa tgtgacaaaa
     1381 caaattgccc gtaatgtctg aaactctgtt ctttcctatg taactgtata cgaaaaattc
     1441 ccccgttaat ttttaatatg tattctgtgt cccatctttg ccccagttta ttttttgtac
     1501 ccctgtgagc gttcaaatta cctcagcatc caacaatctt tccatcttaa catccatccc
     1561 attgtacctc tgaatcttgc ttcgcttacc tcgtaacaaa tatattttta tcggcttaaa
     1621 acctaataaa tcatttacca gaaaaacatt tgtagcaaac aatagaaaaa tccatgaagt
     1681 aaaaaaggag tttaaaaatg aaacagggga tgcatctctg aaccttacaa aaactcgaga
     1741 attcgcagtt gggctgaaca aaaaatcttc aaaaaaaacc cttttataag tctgaaattt
     1801 tcatataacc tgcctcattt gcagatctgc aggaacttgg aatatgatag aaatataaac
     1861 gttatttgtt caagaacact ttcacagttt gttattagat gaataactga gctttcaact
     1921 agtcatagtc aaagaaaata ttagttgggc cctttcgtct cgcgcgtttc ggtgatgacg
     1981 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     2041 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     2101 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     2161 cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc
     2221 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     2281 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     2341 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     2401 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     2461 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     2521 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     2581 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     2641 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     2701 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     2761 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     2821 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     2881 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     2941 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     3001 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     3061 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     3121 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     3181 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     3241 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     3301 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     3361 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     3421 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     3481 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     3541 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     3601 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     3661 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     3721 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     3781 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     3841 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     3901 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     3961 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     4021 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     4081 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     4141 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     4201 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     4261 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     4321 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     4381 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagcttggg
     4441 ctgcaggtcg gctataataa gttcttgaat aaaataattt tcccgacaaa acatgagtat
     4501 ttctttcgaa aataaaagtg caggctaatt agagattatt ctgtaattaa ctgcataatt
     4561 tgtcacgtgc catagtttta cattccacta cgtcatagtt cttaaaatac taatctcctg
     4621 aaaatagaag taggtgaaga aagtttaatt atcagttcta aaatgacaat tgatctttgg
     4681 aatatgttct gaaactaccg atcattgaac agatgctatt tgaatgatat agaattgtat
     4741 atttgcaatt tctgaaacgc gttcttaaag gcacacagat taattcaaaa gggtctggcc
     4801 gcaaaaaggt ttatggtggc cgattttgag ttttgtgtgt gattgctttt tcacaatcag
     4861 tgttttcagg attatgtgat gaactagatc ttcaagtttc gttacatttc atatgttttc
     4921 ggaactcacg aagtacatat tgggtattgt gctcaaaaaa ttcagcaatc agcttcgctc
     4981 cgctgacttt agaacccaaa aaaatagtat ggccaaactg actgtgttac gatcatttca
     5041 atttttcaat acatatttaa gatttctaag agtaagaagg tcaaaaactg ttctggaata
     5101 catatatatt tttcaggtta caattagtca aaaagtgcac tgaaatatac gttttaattt
     5161 cacgaataac ccaattagtt caatgtattt ttggtcaacc aacgttaaag tttggcttcc
     5221 aaccaattat catttctgat caaccacaat gttttttctt tatctgcaag ttaatttttt
     5281 atttttatcc agatgtttgg catatttttc aattcttcac tagcgcccac ttcttgcact
     5341 tccggcgccc tgaatctaat gcatctgttg caagaattga aagaccaatc aacacattgt
     5401 tttcttcacg agatactgaa gaaaatgaat aaaaacagag aaaaagagcc atgtgattag
     5461 tgacaactgt tgctaacaga taccatagct tggacttggt acgtgatggc aacgtatggg
     5521 tcaacaaaaa tgattgcaga gggggtgcaa aacagtcaag tcgagaaaat atgaaaaaca
     5581 gaaaacaaag aacagaaaaa tgggtttgag agtcagtata atttataaaa gaaaaattgt
     5641 acatagaaat taaccatttt tgtagaagaa gttatttttc aagcatcgtt aaaaattatt
     5701 caaagcacct tatttcatat ttaattttaa acatggttaa atgaacaaca cggtgcgcaa
     5761 tcaggaaaac ttgaaatctg aaactgttgt tgtgatcttc ttcgcaactg ttcagatagc
     5821 actagtgtaa tgttaagagt gcgcgaatat aatggaatat aatggatcac acctcctgcc
     5881 atcaggtaaa cgtctctgtt atcacatatt tccaactatt aaatttttac cttttacagt
     5941 tttacatttt tttgaaaaaa gtaacttttt gtcttcaaaa tccctgacga aaatatcaaa
     6001 tattttaatc gagactgcag aggaaccgat tgatgatttg gaaaatccag ctttacctgt
     6061 gtaagaactg aaaagtttca taaccctagg gtattcccag ttacattccc cactggctaa
     6121 caatagcacc cagtttttca tcacctttct tcaaatttct cggcgatttg ttaaaaacaa
     6181 aatttgtgtc ccttctctga tatctctatg tctctaaaca caagttcatc ggaaaacgaa
     6241 ggagggtagg tgttggttgg gctcccgaag tgaaaataga agagcaagaa tagaatatta
     6301 gagagagagt gcagagaggg cgggatagct cccgggattc cgttttcttc ttctttatct
     6361 tcaacgatga tgtgtgtgcg tgttgtatag attctgttgc tcccccacaa ctcgctccga
     6421 aggctcaata caattcaatt gatattggag gagagcctac cggagtggga ggataagaag
     6481 aaacataaga agaagaagaa gaagaagcat gcttctggtt tttgatgcta tgaaaacggc
     6541 acaaaaagat gattgaggtc ccttttcaat accttctctc atctttcaaa tcccattgaa
     6601 acctaaaact tctcaccacg ctttaccatt gttctccaaa aacttatagc aatgtctata
     6661 acttttttat ctctgaaaag cagtgttcca tttttctttt tcctatttta tttcaattgt
     6721 ttctcacatt tcgtttggat tctttgcttg tcaaccagct tcttcttcca cttttaccgt
     6781 ctaattttca gggcagggag ccatcaaacc cacgaccact agatccatct agaggatcct
     6841 atcgatgccc aagaaaaagc gtaaagtccc aaagaagaaa cgaaaggttc ctaaaaagaa
     6901 gcgcaaggtg cccaagaaga aacggaaggt tcccaaaaag aaacgcaaag tttcaaaagg
     6961 agaggaattg ttcaccggcg tcgttccaat cctcgttgaa ctcgacggcg acgtcaacgg
     7021 gcacaagttt tccgtctctg gagagggaga gggagacgca acctacggca aattgaccct
     7081 taaacttatt tgcaccacag gtaagtttcg atttttgcta tctttttcac gatttttact
     7141 ctctttttct actaactaac ctcctttttg aacatttttt caggaaaatt gccagttcca
     7201 tggccaacat tggtcacaac cttgggatac ggcttgcaat gcttcgcacg ttaccccgat
     7261 cacatgaaac aacacgactt ttttaaatcc gcaatgccag aaggatacgt acaagaacgc
     7321 acaattttct tcaaagatga cggaaactac aagacaaggt aagtttttcc caactttttc
     7381 ggactttttg accgcttttt atactaacta acggactttt tcgtcctttt tcagggccga
     7441 agtcaagttt gaaggagaca ccctcgtcaa ccgcatcgaa cttaaaggaa ttgacttcaa
     7501 agaagacgga aacattctcg gacacaaact cgaatacaac tacaattctc acaacgttta
     7561 cataacagcc gacaaacaaa agaacggaat caaggtaagt ttcgcttttt gatgattttt
     7621 ggaccatttt taggaatttt tcgatactaa ctaactattt ttgcctgctt tttcaggcaa
     7681 acttcaagat tcgtcacaac attgaggacg gaggagtcca acttgctgac cattaccagc
     7741 aaaacacccc gattggcgac ggacccgtcc tccttccgga caaccactac ctctcatacc
     7801 agagcgccct ctcgaaagac ccgaacgaga agaggtaagt ttatcgtttt tcgacgattt
     7861 ttacccattt ttacgccatt tttgatacta actaacgcct ttttaccgat ttttcaggga
     7921 ccacatggtc cttttggaat ttgttacggc cgcaggcatc acccacggca tggacgaact
     7981 ctacaaatag cattcgtaga attccaactg agcgccggtc gctaccatta ccaacttgtc
     8041 tggtgtcaaa aataatagg
//
