LOCUS       sequence-12967-e        4583 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 4583)
  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 4583)
  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: pHZ149
FEATURES             Location/Qualifiers
     source          1..4583
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             585..605
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             630..650
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(2023..2045)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2145..2164
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2242..2260)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2328..2432
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2433..2501,2502..3293)
                     /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(2651..2670)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3464..4052
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3953..3972
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4206..4223
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4340..4361
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4376..4381,4382..4399,4400..4406)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4414..4430
                     /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     4419..4441
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4438..4454
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4438..4454
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttgaattgg gtgataaaat gcttgaattg ggtgataaaa
       61 tgcttgaatt gggtgataaa atgcttggtc ccttgaattg ggtgataaaa tgcttgaatt
      121 gggtgataaa atgcttgaat tgggtgataa aatgcttggt cccttgaatt gggtgataaa
      181 atgcttgaat tgggtgataa aatgcttgaa ttgggtgata aaatgcttgg tcccttgaat
      241 tgggtgataa aatgcttgaa ttgggtgata aaatgcttga attgggtgat aaaatgcttg
      301 gtcgactcta gacacttttc agctacctag atacatggat atccccgcct cccaatccac
      361 ccacccaggg aaaaagaagg gctcgccgaa aaatcaaagt tatctccagg ctcgcgcatc
      421 ccaccgagcg gttgacttct ctccaccact tttcatttta accctcgggg tacgggattg
      481 gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga
      541 ggacccttgc ttggagggta ccgagctcag aaaaaatgac tgcaccgaaa aagaaacgca
      601 aagtaccgag ctcagaaaaa atgaccgctc caaagaagaa acgcaaagta ccggtcgcca
      661 ccatgaggtc ttccaagaat gttatcaagg agttcatgag gtttaaggtt cgcatggaag
      721 gaacggtcaa tgggcacgag tttgaaatag aaggcgaagg agaggggagg ccatacgaag
      781 gccacaatac cgtaaagctt aaggtaacca aggggggacc tttgccattt gcttgggata
      841 ttttgtcacc acaatttcag tatggaagca aggtatatgt caagcaccct gccgacatac
      901 cagactataa aaagctgtca tttcctgaag gatttgtaag tttaaacggg gtgttactaa
      961 ctaacaaata acatttaaat tttcagaaat gggaaagggt catgaacttt gaagacggtg
     1021 gcgtcgctac tgtcacccaa gattccagct tgcaagatgg ctgtttcatc tacaaggtca
     1081 agttcattgg cgtgaacttt ccttccgatg gacctgttat gcaaaagaag acaatgggct
     1141 gggaagccag cactgagcgt ttgtatcctc gtgatggcgt gttgaaagga gagattcata
     1201 aggctctgaa gctgaaagac gggtaagttt aaactatatc ttactaacta actgtagaga
     1261 tttaaatttt cagtggtcat tacctagttg aattcaaaag tatttacatg gcaaagaagc
     1321 ctgtgcagct accagggtac tactatgttg actccaaact ggatataaca agccacaacg
     1381 aagactatac aatcgttgag cagtatgaaa gaaccgaggg acgccaccat ctgttccttt
     1441 agcggccgcg actctagaat tccaactgag cgccggtcgc taccattacc aacttgtctg
     1501 gtgtcaaaaa taataggcct actagccggc catacaagta atccggatga tcgacgccaa
     1561 cgtcgttgaa ttttcaaatt ttaaatactg aatatttgtt ttttttccta ttatttattt
     1621 attctctttg tgtttttttt cttgctttct aaaaaattaa ttcaatccaa atctaaacat
     1681 gagcggtttt ttttctcttt ccgtctccca attcgtattc cgctcctctc atctgaacac
     1741 aatgtgcaag tttatttatc ttctcgcttt catttcatta ggacgtgggg ggaattggtg
     1801 gaagggggaa acacacaaaa ggatgatgga aatgaaataa ggacacacaa tatgcaacaa
     1861 cattcaattc agaaatatgg aggaaggttt aaaagaaaac ataaaaatat atagaggagg
     1921 aaggaaaact agtaaaaaat aagcaaagaa attaggcgaa cgatgagaat tgtcctcgct
     1981 tgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac acatgcagct
     2041 cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag cccgtcaggg
     2101 cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat cagagcagat
     2161 tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa ggagaaaata
     2221 ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt aatgtcatga
     2281 taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta
     2341 tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa taaccctgat
     2401 aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc
     2461 ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa acgctggtga
     2521 aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa ctggatctca
     2581 acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg atgagcactt
     2641 ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa gagcaactcg
     2701 gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc acagaaaagc
     2761 atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc atgagtgata
     2821 acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta accgcttttt
     2881 tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag
     2941 ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca acgttgcgca
     3001 aactattaac tggcgaacta cttactctag cttcccggca acaattaata gactggatgg
     3061 aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc tggtttattg
     3121 ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca ctggggccag
     3181 atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca actatggatg
     3241 aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg taactgtcag
     3301 accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa tttaaaagga
     3361 tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt gagttttcgt
     3421 tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat cctttttttc
     3481 tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg gtttgtttgc
     3541 cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga gcgcagatac
     3601 caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac tctgtagcac
     3661 cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt ggcgataagt
     3721 cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag cggtcgggct
     3781 gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc gaactgagat
     3841 acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag gcggacaggt
     3901 atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca gggggaaacg
     3961 cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt cgatttttgt
     4021 gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc tttttacggt
     4081 tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc cctgattctg
     4141 tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg
     4201 agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc
     4261 ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac tggaaagcgg
     4321 gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc caggctttac
     4381 actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa tttcacacag
     4441 gaaacagcta tgaccatgat tacgccaagc tgtaagttta aacatgatct tactaactaa
     4501 ctattctcat ttaaattttc agagcttaaa aatggctgaa atcactcaca acgatggata
     4561 cgctaacaac ttggaaatga aat
//
