LOCUS       sequence-217-exp        4895 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 4895)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4895)
  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: L2531
FEATURES             Location/Qualifiers
     source          1..4895
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2335..2357)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2457..2476
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2554..2572)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2640..2744
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2745..2813,2814..3605)
                     /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(2963..2982)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3776..4364
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4265..4284
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4518..4535
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4652..4673
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4688..4693,4694..4711,4712..4718)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4726..4742
                     /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     4731..4753
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4750..4766
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4750..4766
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccag cttgcatgcc tgcaggtcga
       61 ggcatttgaa ttgggggtgg tggacagtaa ctgtctgtaa taataattac tcctgaccag
      121 gttgcaattc gagttttgat aagcataatt ataccttgta cattgtgggt tttgtgctgt
      181 ggacgtttta ttgtgacatc cccataagct acaagaacaa aaatgaaatg taaaagtatt
      241 gaaaacgtcg tacattttat atctgagtag tatcctttgc tttaaatgtc cataaaacta
      301 attttataat caataaaaca acgtttgtaa atcaactgag tttacaagta gagacattga
      361 gggatacttt cactatgcta aagtgataat cgaccaaata ataacttcac tttggtattt
      421 attcctgtct tataaatgtt atgtatgaat taaattcata tgcatatggc tcactctgac
      481 aatttttaat aatcttccag atcaatattg actaccgatg cgggtggtct tttgctttga
      541 attctgctga actttacacc ccgaacagca atgtgtgctt cagcctaaaa aaaagtaagt
      601 gtgttaatca gtgcccccga ttcttcattt tttgcccctc tctcccgttt cgtcggcaaa
      661 agaagagaaa ataaagataa gtctcaagat aggttggtaa tcgctaaagt ggttgtgtgg
      721 ataagagtag caaaatggca ggaagagcac tttgcgcgca cacactgtac tcattgttct
      781 ggataaaatt ctctcgttgt ttgccgtcgg atgtctgcct ctctgcattg agccggcttc
      841 ttcactatct ttagttaacc taaaatgccg tttcttttct cgtatcccca ctatcccgtt
      901 gaggttctct gctctcttcg ctccctaccg ccagcgagca actatccgtg ggggcgcctt
      961 gctcggaaga tgggggggaa gaaagaagat ttttgctatt tgcacttgag aaagagactt
     1021 ttcctgcgtc gatggttaga gaacagtgtg cagacacttt tcagctacct agaattacaa
     1081 ttggatatcc ccgcctccca atccacccac ccagggaaaa agaagggctc gccgaaaatc
     1141 aaagttatct ccaggctcgc gcatcccacc gagcggttga cttctctcca ccacttttca
     1201 ttttaaccct cggggtacgg gattggccaa aggacccaaa ggtatgtttc gaatgatact
     1261 aacataacat agaacatttt caggaggacc cttggctagc gtcgacggta ccggtcagat
     1321 atcactcgag gagctcccga gatcttcagc gctcagcatt cgtagaattg taagtttaaa
     1381 ctttctcata ctaactaacg cgctctattt aaattttcag aattccaact gagcgccggt
     1441 cgctaccatt accaacttgt ctggtgtcaa aaataatagg ggccgctgtc atcagagtaa
     1501 gtttaaactg agttctacta actaacgagt aatatttaaa ttttcagcat ctcgcgcccg
     1561 tgcctctgac ttctaagtcc aattactctt caacatccct acatgctctt tctccctgtg
     1621 ctcccacccc ctatttttgt tattatcaaa aaacttctct taatttcttt gttttttagc
     1681 ttcttttaag tcacctctaa caatgaaatt gtgtagattc aaaaatagaa ttaattcgta
     1741 ataaaaagtc gaaaaaaatt gtgctccctc cccccattaa taataattct atcccaaaat
     1801 ctacacaatg ttctgtgtac acttcttatg ttttttactt ctgataaatt tttttgaaac
     1861 atcatagaaa aaaccgcaca caaaatacct tatcatatgt tacgtttcag tttatgaccg
     1921 caatttttat ttcttcgcac gtctgggcct ctcatgacgt caaatcatgc tcatcgtgaa
     1981 aaagttttgg agtatttttg gaatttttca atcaagtgaa agtttatgaa attaattttc
     2041 ctgcttttgc tttttggggt ttcccctatt gtttgtcaag atttcgagga cggcgttttt
     2101 cttgctaaaa tcacaagtat tgatgagcac gatgcaagaa agatcggaag aaggtttggg
     2161 tttgaggctc agtggaaggt gagtagaagt tgataatttg aaagtggagt agtgtctatg
     2221 gggtttttgc cttaaatgac agaatacatt cccaatatac caaacataac tgtttcctac
     2281 tagtcggccg tacgggccct ttcgtctcgc gcgtttcggt gatgacggtg aaaacctctg
     2341 acacatgcag ctcccggaga cggtcacagc ttgtctgtaa gcggatgccg ggagcagaca
     2401 agcccgtcag ggcgcgtcag cgggtgttgg cgggtgtcgg ggctggctta actatgcggc
     2461 atcagagcag attgtactga gagtgcacca tatgcggtgt gaaataccgc acagatgcgt
     2521 aaggagaaaa taccgcatca ggcggcctta agggcctcgt gatacgccta tttttatagg
     2581 ttaatgtcat gataataatg gtttcttaga cgtcaggtgg cacttttcgg ggaaatgtgc
     2641 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     2701 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     2761 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     2821 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     2881 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     2941 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     3001 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     3061 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     3121 ccatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     3181 taaccgcttt tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     3241 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     3301 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     3361 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     3421 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     3481 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg
     3541 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     3601 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     3661 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     3721 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     3781 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     3841 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     3901 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     3961 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     4021 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     4081 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     4141 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     4201 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
     4261 cagggggaaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
     4321 gtcgattttt gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg
     4381 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
     4441 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
     4501 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
     4561 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     4621 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttagctcact cattaggcac
     4681 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     4741 aatttcacac aggaaacagc tatgaccatg attacgccaa gctgtaagtt taaacatgat
     4801 cttactaact aactattctc atttaaattt tcagagctta aaaatggctg aaatcactca
     4861 caacgatgga tacgctaaca acttggaaat gaaat
//
