LOCUS       sequence-12964-e        6257 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 6257)
  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 6257)
  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: pHZ081
FEATURES             Location/Qualifiers
     source          1..6257
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3832..3854)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3954..3973
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4051..4069)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4137..4241
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4242..4310,4311..5102)
                     /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(4460..4479)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5273..5861
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5762..5781
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6015..6032
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6149..6170
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6185..6190,6191..6208,6209..6215)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6223..6239
                     /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     6228..6250
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
ORIGIN
        1 atgaccatga ttacgccaag cttcagtgct gaaaaaattc aagaactctg ttcaaagaaa
       61 taattacatc taattaaatt ttatttgtct tcttataaat aaattgttgt atatatctca
      121 agccttttgt tgatatttaa tccacagacc atgatattga tcatctatta attcttcgaa
      181 catttccatt cagaacactt ttgtaacaat ttaaatcgtt ttgaaatgaa ttcgatgtgt
      241 ttatatttta atcaggaggc gtttggattt ataaacgagc gtccgacagg attagtgtgc
      301 cactttcttt ttcagtccac acttgtcccc tctcaacact tttttgttat tctctcactt
      361 tatcctcacc tctcgcctaa agttcatttc aaatttttga ttcatttctt actgtgtgaa
      421 atcaagaact agacggagtc ggagtggttg gctcgagatg cttcaatgag tcgacggact
      481 gcgtcgaagt tactagtttt tgtattttta tgctctttat gttttggaac tcaacggtat
      541 gatatgccga gaaaaattga cttgtttaat gatttgatta cacaatcgac aactccagca
      601 tctccaaagg tgaataatca aatatacaac tttcaaacca aagcatttat aatttttgta
      661 gtttgtttta gtttttaatt ttcaagtttt cttttgataa ttgttattct aaaaaattga
      721 aagcaatggg tagttttgtt caaagttcaa atttatacaa caacgtctac ctactaaaaa
      781 atggtatatg ccgagtttta agttgattcg aaagttgatc tcgtataaac tagctgatct
      841 gaacttccga actcaaagtt tttgccgaat tagcacgaaa cacaagttcc caaaatattt
      901 caaatgtttt cagtgccaat gtcttccacc aacaactcca tccacccctc caaactgtat
      961 tccttacgat tctcgattgc aagcagcaag tcttgaagaa gctattgtgg cattcccaga
     1021 tcttaccatt actcgacaag agaagacaca acaatctgta attctttttt tttcattatg
     1081 ttttcggatc tcaattaact ttcaaattat ggaaatttgt tttaaagctt tttcggagtg
     1141 taaagtaact ataatatttc agactgcaac actaaacaac tgcaaaacta aacaatgccg
     1201 tgattgctat aaagatctca gaagccagtt gcgaaaagta ggaagcctct tgcagaatcg
     1261 cttctagaaa tgttcacttc caattttcag gtcggtcttc ttccaggaac cattgaccag
     1321 gtgtttcaca atcagcggaa cttcacaact tgtcaaaaat atagatttgc aagacaggat
     1381 aaaggagtct atgagaagaa aaagaaagcc aaacaacatt atgattggga ttatgtagaa
     1441 tgtaagagaa cattcagtgt tctttttttg gaaaaactct caagtacaaa ttgaaaggtt
     1501 tcagatgatg aagatgaaga tgatgattac ttttgggatg gattattttg gaagaagaag
     1561 agaaatgtat taaagaaaat tgtaaaacga gacgttgaag ctactactgc aatttcacaa
     1621 ccacctaaca gtacagcaat gaactcaacc ggaattatag gtttgtattg aagacataat
     1681 tttttttcta aaaacttctt attggtatat atttttcaac actctgaaaa taagatatat
     1741 tcaaaacaat aattttaagt gaccgtcata aataattatg aaaagaaaac aaccattcaa
     1801 aggttttttt aaaaattttt actcaaaatg cgtgtattgt tctccctcaa aaaattattt
     1861 ccgggtcgtg aattccctta cgagaatttc aattcccctc gggtggtcat catctcaaat
     1921 tccaatcttc tttttcattt tcaatctcta acaatttcaa attactagaa cccacaaacc
     1981 aatgttcgat aacggcatgc aaccttgaga gagtctccga tttgatgtgt aaccttcatt
     2041 aaaccctgta atacaagtca tgtcgacaaa ccagatcttc atcccttcaa catttggcag
     2101 aaaagtgcaa ttcgtctttt acaggaattc gatttccgat ctcttgcaca actcgaggcg
     2161 tcactccaga tggattagga actgtttccc tttgtagtac atgttgggtg tggcgacgtc
     2221 ttccgagtac ctactatcct gcatatctta acgaagtcgt gtgcgattat gcggacacat
     2281 cgtgtctgag tggtagtgaa aagacagaga aagtggggga aaaagaaaat accatttttt
     2341 caggatacgc gtcatgccaa actggaaccc aacagttgaa tgtgctacga aatgacagcg
     2401 ggaaattgat tccaatatcg gtatcagctg gaatcaattg tgaatgtaga ttagcagttg
     2461 gatcaacact tgaatcactc gtacttggac aaggaatttc caaagcaatg ccgccaattg
     2521 acacaactag tacaaaacca ccaaatttgg caacaagcac aacaagccat tcctaattta
     2581 tttgggtatt ttactcttga taattgtatt aatgatcatt taatgttgtt gaatatcaca
     2641 catttaacat attcaatcga agaaatataa cgacgttcca caacatccag tatacgtctt
     2701 cctctgtatt gagttttatt ctttgacatt gtgtcatgtt tcagaacata gaaatatacc
     2761 aaagtgttga gaagcattgt ataattcaat tgaatctttt tcagtttacc tcttctcctt
     2821 tttcaccgtt tccgtcatca aaacgccgtt attcaaggtg agattatggc gataaatgaa
     2881 tgccattcac aaaattgcaa gacactatta aagaagaaga ggtttagaga tttgtaccga
     2941 tcatagcttc ttctgagaaa gcgaccttgg cggccgtact tgaagagaag caatttttat
     3001 cgccgtagaa gatgttgcct gccacccacc ctgtacgttg tttttttgct tatcaacatc
     3061 tgaattttgt tgtcttttgc accgcctact tttactatgg aatttgagga aatataatat
     3121 gaattgaaga gaatacgcta cctgtcaaag aaacgtcacg gttagggtag tataaaaggg
     3181 aagggtaaca ctgaataagg tacctggctt tcagtcatag agggatccaa aaaaaatgct
     3241 tccggcctgt tccctgagac ctcaagtgtg agtgtactat tgatgcttca cacctgggct
     3301 ctccgggtac caggacggtt tgagcagatg ctagccggcc atacaagtaa tccggatgat
     3361 cgacgccaac gtcgttgaat tttcaaattt taaatactga atatttgttt tttttcctat
     3421 tatttattta ttctctttgt gttttttttc ttgctttcta aaaaattaat tcaatccaaa
     3481 tctaaacatg agcggttttt tttctctttc cgtctcccaa ttcgtattcc gctcctctca
     3541 tctgaacaca atgtgcaagt ttatttatct tctcgctttc atttcattag gacgtggggg
     3601 gaattggtgg aagggggaaa cacacaaaag gatgatggaa atgaaataag gacacacaat
     3661 atgcaacaac attcaattca gaaatatgga ggaaggttta aaagaaaaca taaaaatata
     3721 tagaggagga aggaaaacta gtaaaaaata agcaaagaaa ttaggcgaac gatgagaatt
     3781 gtcctcgctt gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     3841 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     3901 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     3961 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     4021 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     4081 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4141 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4201 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     4261 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     4321 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     4381 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     4441 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     4501 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     4561 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     4621 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     4681 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     4741 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     4801 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     4861 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     4921 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     4981 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5041 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5101 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5161 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     5221 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     5281 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     5341 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     5401 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     5461 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     5521 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     5581 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     5641 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     5701 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     5761 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     5821 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     5881 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     5941 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6001 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6061 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6121 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6181 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     6241 ttcacacagg aaacagc
//
