LOCUS       sequence-12966-e        4168 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 4168)
  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 4168)
  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: pHZ146
FEATURES             Location/Qualifiers
     source          1..4168
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1297..1319)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     1419..1438
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(1516..1534)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1602..1706
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1707..1775,1776..2567)
                     /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(1925..1944)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2738..3326
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3227..3246
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3480..3497
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3614..3635
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3650..3655,3656..3673,3674..3680)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3688..3704
                     /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     3693..3715
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3712..3728
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3712..3728
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 acacttttca gctacctaga tacatggata tccccgcctc ccaatccacc cacccaggga
       61 aaaagaaggg ctcgccgaaa aatcaaagtt atctccaggc tcgcgcatcc caccgagcgg
      121 ttgacttctc tccaccactt ttcattttaa ccctcggggt acgggattgg ccaaaggacc
      181 caaaggtatg tttcgaatga tactaacata acatagaaca ttttcaggag gacccttggc
      241 tagcatgctt ccggcctgtt ccctgagacc tcaagtgtga gtgtactatt gatgcttcac
      301 acctgggctc tccgggtacc aggacggttt gagcagatct ttttttctag cattctcatg
      361 gagccgcatt cgtagaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt
      421 gtcaaaaata ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa
      481 cgagtaatat ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta
      541 ctcttcaaca tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta
      601 tcaaaaaaac ttcttcttaa tttctttgtt ttttagcttc ttttaagtca cctctaacaa
      661 tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa aaaaattgtg
      721 ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc tgtgtacact
      781 tcttatgttt tttttacttc tgataaattt tttttgaaac atcatagaaa aaaccgcaca
      841 caaaatacct tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac
      901 gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg
      961 gaatttttca atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggg
     1021 tttcccctat tgtttgtcaa gagtttcgag gacggcgttt ttcttgctaa aatcacaagt
     1081 attgatgagc acgatgcaag aaagatcgga agaaggtttg ggtttgaggc tcagtggaag
     1141 gtgagtagaa gttgataatt tgaaagtgga gtagtgtcta tggggttttt gccttaaatg
     1201 acagaataca ttcccaatat accaaacata actgtttcct actagtcggc cgtacgggcc
     1261 ctttcgtctc gcgcgtttcg gtgatgacgg tgaaaacctc tgacacatgc agctcccgga
     1321 gacggtcaca gcttgtctgt aagcggatgc cgggagcaga caagcccgtc agggcgcgtc
     1381 agcgggtgtt ggcgggtgtc ggggctggct taactatgcg gcatcagagc agattgtact
     1441 gagagtgcac catatgcggt gtgaaatacc gcacagatgc gtaaggagaa aataccgcat
     1501 caggcggcct taagggcctc gtgatacgcc tatttttata ggttaatgtc atgataataa
     1561 tggtttctta gacgtcaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
     1621 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
     1681 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
     1741 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
     1801 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
     1861 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
     1921 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
     1981 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
     2041 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
     2101 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
     2161 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
     2221 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
     2281 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
     2341 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
     2401 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
     2461 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
     2521 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     2581 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     2641 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     2701 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     2761 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     2821 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     2881 ctgtccttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     2941 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     3001 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     3061 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     3121 agcgtgagca ttgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     3181 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     3241 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     3301 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     3361 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     3421 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     3481 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     3541 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     3601 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     3661 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     3721 gctatgacca tgattacgcc aagctgtaag tttaaacatg atcttactaa ctaactattc
     3781 tcatttaaat tttcagagct taaaaatggc tgaaatcact cacaacgatg gatacgctaa
     3841 caacttggaa atgaaataag cttgcatgcc tgcaggcctt gaattgggtg ataaaatgct
     3901 tgaattgggt gataaaatgc ttgaattggg tgataaaatg cttggtccct tgaattgggt
     3961 gataaaatgc ttgaattggg tgataaaatg cttgaattgg gtgataaaat gcttggtccc
     4021 ttgaattggg tgataaaatg cttgaattgg gtgataaaat gcttgaattg ggtgataaaa
     4081 tgcttggtcc cttgaattgg gtgataaaat gcttgaattg ggtgataaaa tgcttgaatt
     4141 gggtgataaa atgcttggtc gactctag
//
