LOCUS       sequence-12965-e        5065 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 5065)
  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 5065)
  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: pHZ126
FEATURES             Location/Qualifiers
     source          1..5065
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1457..1477
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2506..2528)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2628..2647
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2725..2743)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2811..2915
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2916..2984,2985..3776)
                     /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(3134..3153)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3947..4535
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4436..4455
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4689..4706
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4823..4844
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4859..4864,4865..4882,4883..4889)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4897..4913
                     /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     4902..4924
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4921..4937
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4921..4937
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgaaa tcacaaattc catttttatg tcatacttca gatttaaaca aatttatttt
       61 atgtgtgttt ttttaatggt atgtcctaac gattttccta attgacaact attatagatt
      121 gaaaacacag aatgccaagt aacgtaagaa attttttttt cgaataccag acataatttc
      181 aataaaagaa aaatctttaa aaaaagttta actttatact acaataattt tggtttttat
      241 gaaggaatct gtattgcggc acatcatgta tttgctcaag gtttcgttgg ttgacatatt
      301 tttgtgatcc gtggaatgag cttgcataga gtttagtcat aattgggttt attttacata
      361 cagtttgaag tatgttttgt ctgttattcg aaatttttta gttagtttgg taagttgaaa
      421 agaatttaca aaatttaact aaacgaggac gctcatctcg attcttataa ttccataata
      481 attctacggt aaaagtcaag ttatgcctca aaacatgtaa tacataatta ccaaaactac
      541 ttaattaccg catttcctta gtattttaat agtggtgtag tcgaattttt tttattgctt
      601 tattagactc aaaattgtct gcaaacaccc aatttcataa tgaaacttat tgaaaacaat
      661 acactttgaa acaatttagt atttcaggaa taaggtcact ggaacttcga aaatgataat
      721 ttgaataacc gttaataaaa tattcaaacc aatttgcata atcttgattt tgtatcatga
      781 tggtatagaa tgggactttt gaaagatgtg aagtttcaat tcagtaagta caactttaaa
      841 attgggctgc aagatttctt cttaaatttt caacagtttc aaaacactga aatcatgctg
      901 aactatggtt agagactcga caactttcct aaattttgga gcagtttcaa tggttttcga
      961 atgtatacta catttgtacc attttatgtg ttagtgtagg ttgctttgct tctaccttca
     1021 ctctcaaatt cttttttccg cggtaatttt tcaactatcg aaacaaagaa gaagtcatga
     1081 ataactgtcc gcatagagtt ctcacccgtc ccctttcttt tcaccgagca gtcacgagag
     1141 aaagaagaag aagcggaccg ctgcagagat tcttcgtagc ggagccccgc ccatcctctg
     1201 ttacgcatat gtttggatct catttcttcc actcattaac gttccaggta tgtttttttt
     1261 gttgattttt taatgtgtta cttttcatat gtttattact tgaactgacc gatttcaaat
     1321 atgattttca cagatgtcta gaggatcccc gggattggcc aaaggaccca aaggtatgtt
     1381 tcgaatgata ctaacataac atagaacatt ttcaggagga cccttgaggg taccgagctc
     1441 agaaaaaatg actgctccaa agaagaagcg taaggtaccg gtagaaaaaa tgcttccggc
     1501 ctgttccctg agacctcaag tgtgagtgta ctattgatgc ttcacacctg ggctctccgg
     1561 gtaccaggac ggtttgagca gatgaattcc aactgagcgc cggtcgctac cattaccaac
     1621 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     1681 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     1741 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     1801 ttgttattat caaaaaaact tcttcttaat ttctttgttt tttagcttct tttaagtcac
     1861 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     1921 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     1981 gtgtacactt cttatgtttt ttttacttct gataaatttt ttttgaaaca tcatagaaaa
     2041 aaccgcacac aaaatacctt atcatatgtt acgtttcagt ttatgaccgc aatttttatt
     2101 tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct catcgtgaaa aagttttgga
     2161 gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa ttaattttcc tgcttttgct
     2221 ttttgggggt ttcccctatt gtttgtcaag agtttcgagg acggcgtttt tcttgctaaa
     2281 atcacaagta ttgatgagca cgatgcaaga aagatcggaa gaaggtttgg gtttgaggct
     2341 cagtggaagg tgagtagaag ttgataattt gaaagtggag tagtgtctat ggggtttttg
     2401 ccttaaatga cagaatacat tcccaatata ccaaacataa ctgtttccta ctagtcggcc
     2461 gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt gaaaacctct gacacatgca
     2521 gctcccggag acggtcacag cttgtctgta agcggatgcc gggagcagac aagcccgtca
     2581 gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt aactatgcgg catcagagca
     2641 gattgtactg agagtgcacc atatgcggtg tgaaataccg cacagatgcg taaggagaaa
     2701 ataccgcatc aggcggcctt aagggcctcg tgatacgcct atttttatag gttaatgtca
     2761 tgataataat ggtttcttag acgtcaggtg gcacttttcg gggaaatgtg cgcggaaccc
     2821 ctatttgttt atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct
     2881 gataaatgct tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg
     2941 cccttattcc cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg
     3001 tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc
     3061 tcaacagcgg taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca
     3121 cttttaaagt tctgctatgt ggcgcggtat tatcccgtat tgacgccggg caagagcaac
     3181 tcggtcgccg catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa
     3241 agcatcttac ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg
     3301 ataacactgc ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt
     3361 ttttgcacaa catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg
     3421 aagccatacc aaacgacgag cgtgacacca cgatgcctgt agcaatggca acaacgttgc
     3481 gcaaactatt aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga
     3541 tggaggcgga taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta
     3601 ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc
     3661 cagatggtaa gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg
     3721 atgaacgaaa tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt
     3781 cagaccaagt ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa
     3841 ggatctaggt gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     3901 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt
     3961 ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt
     4021 tgccggatca agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga
     4081 taccaaatac tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag
     4141 caccgcctac atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata
     4201 agtcgtgtct taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg
     4261 gctgaacggg gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga
     4321 gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca
     4381 ggtatccggt aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa
     4441 acgcctggta tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt
     4501 tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac
     4561 ggttcctggc cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt
     4621 ctgtggataa ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga
     4681 ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg cccaatacgc aaaccgcctc
     4741 tccccgcgcg ttggccgatt cattaatgca gctggcacga caggtttccc gactggaaag
     4801 cgggcagtga gcgcaacgca attaatgtga gttagctcac tcattaggca ccccaggctt
     4861 tacactttat gcttccggct cgtatgttgt gtggaattgt gagcggataa caatttcaca
     4921 caggaaacag ctatgaccat gattacgcca agctgtaagt ttaaacatga tcttactaac
     4981 taactattct catttaaatt ttcagagctt aaaaatggct gaaatcactc acaacgatgg
     5041 atacgctaac aacttggaaa tgaaa
//
