LOCUS       sequence-258-exp        4669 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 4669)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4669)
  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"
COMMENT     Sequence Label: L4404
FEATURES             Location/Qualifiers
     source          1..4669
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     598..614
                     /label=pBluescriptKS
                     /note="For pBluescript vector"
     primer_bind     599..615
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1824..1843)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(1825..1843)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(1853..1870)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1853..1869)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1862..1884)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2011..2466
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2098..2117)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2308..2329
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        2492..2596
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2597..2665,2666..3457)
                     /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(2815..2834)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3628..4216
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4117..4136
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4370..4387
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4504..4525
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4540..4545,4546..4563,4564..4570)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4578..4594
                     /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     4583..4605
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4602..4618
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4602..4618
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     4637..4657
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        4639..4657
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 gagctccacc gcggtggcgg ccccccctcg agttgagaag taagctggga cttcagtctg
       61 gtgagttggt tgacttggga ttcggcatct tcgagttgac ggacaaggtc tccattctca
      121 gagtgaaggc gtcccttaag ggaagtgaag tcttggagtt ggcgagattg ctcatcagcc
      181 ttagattgaa gctcggtgag ttgaagttcg aactgcttgg caagcttttc gttgttgagc
      241 tttccagagg tttcttggtc aagttgagca gcaaggtcct cagcgtcacg aacagcttga
      301 gccttgtcct tctcgacctt agccttggcc ttgttgagtt gatcgagttg gtcggtgagc
      361 tcagcgacag cgtcggtgtg cttcttgcga agtccaccga gttggttctc gtggttcatg
      421 ttggcctcct ccaagtctct gcggagcttg gcaagttcag cctcacgctt cttgttgacc
      481 tcaacctggg cggcagtagc tccaccttgc tcgtcaagct tctcaccgag ctcttccaac
      541 tcgcgttgga ggtcgctctt ggcacggtca gccttggaac gggattgacg ttcattctcg
      601 aggtcgacgg tatcgataag cttgatatcg aattcctgca gtatttaaaa atcgaaacta
      661 gaatgtttcc tatttcagac ctactccgga cttttctgcg ttgtcatcaa cccatacaag
      721 cgtctcccaa tctacaccga ctcttgcgcc cgtatgttca tgggaaagag aaagacagaa
      781 atgccaccac atttgttcgc tgtctctgac gaagcctacc gtaacatgct ccaagaccac
      841 gagaaccagt ctatgctcat taccggagaa tctggagccg gaaagactga gaacaccaag
      901 aaggttattt gctacttcgc cgctgtcggt gcctcccaac aggaaggtgg agctgaagtt
      961 gatccaaata agaagaaggt cacccttgag gaccaaatcg tccagaccaa tccagtactt
     1021 gaagccttcg gtaacgccaa gactgtccgt aacaacaact catcccgttt cggaaagttc
     1081 atccgtatcc acttcaacaa gcacggacgc ctcgcatctt gcgatatcga gcattgtgag
     1141 ttactttttg atttatattc tagcatagct caataaaaat tatgcaacta ttgtttaaac
     1201 tataaatttt gcagacttgc tcgacctcga gaatgaacgt caatcccgtt ccaaggctga
     1261 ccgtgccaag agcgacctcc aacgcgagtt ggaagagctc ggtgagaagc ttgacgagca
     1321 aggtggagct actgccgccc aggttgaggt caacaagaag cgtgaggctg aacttgccaa
     1381 gctccgcaga gacttggagg aggccaacat gaaccacgag aaccaactcg gtggacttcg
     1441 caagaagcac accgacgctg tcgctgagct caccgaccaa ctcgatcaac tcaacaaggc
     1501 caaggctaag gtcgagaagg acaaggctca agctgttcgt gacgctgagg accttgctgc
     1561 tcaacttgac caagaaacct ctggaaagct caacaacgaa aagcttgcca agcagttcga
     1621 acttcaactc accgagcttc aatctaaggc tgatgagcaa tctcgccaac tccaagactt
     1681 cacttccctt aagggacgcc ttcactctga gaatggagac cttgtccgtc aactcgaaga
     1741 tgccgaatcc caagtcaacc aactcaccag actgaagtcc cagcttactt ctcaactcga
     1801 gggggggccc ggtacccaat tcgccctata gtgagtcgta ttacgcgcgc tcactggccg
     1861 tcgttttaca acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag
     1921 cacatccccc tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc
     1981 aacagttgcg cagcctgaat ggcgaatggg acgcgccctg tagcggcgca ttaagcgcgg
     2041 cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta gcgcccgctc
     2101 ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt caagctctaa
     2161 atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac cccaaaaaac
     2221 ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt tttcgccctt
     2281 tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga acaacactca
     2341 accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg gcctattggt
     2401 taaaaaatga gctgatttaa caaaaattta acgcgaattt taacaaaata ttaacgctta
     2461 caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta
     2521 aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata
     2581 ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc
     2641 ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga
     2701 agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct
     2761 tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg
     2821 tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta
     2881 ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat
     2941 gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt
     3001 acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga
     3061 tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga
     3121 gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga
     3181 actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc
     3241 aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc
     3301 cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg
     3361 tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat
     3421 cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata
     3481 tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct
     3541 ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga
     3601 ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg
     3661 cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc
     3721 aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgtccttct
     3781 agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc
     3841 tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt
     3901 ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg
     3961 cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct
     4021 atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag
     4081 ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag
     4141 tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg
     4201 gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg
     4261 gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac
     4321 cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt
     4381 gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     4441 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     4501 aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc
     4561 tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca
     4621 tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctg
//
