LOCUS       sequence-244-exp        4745 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 4745)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 4745)
  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: L4058
FEATURES             Location/Qualifiers
     source          1..4745
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(422..450)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1198..1218
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(2185..2207)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2307..2326
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2404..2422)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2490..2594
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2595..2663,2664..3455)
                     /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(2813..2832)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3626..4214
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4115..4134
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4368..4385
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4502..4523
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4538..4543,4544..4561,4562..4568)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4576..4592
                     /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     4581..4603
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     4600..4616
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     4600..4616
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccgg gattggccaa aggacccaaa
       61 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      121 accgccggca atgccacctt caacatcatt gctgctcctc gcagcacttc ttccattcgc
      181 tttaccagca agcgattgga agactggaga agtcactccg gataatgcaa cagtctgggt
      241 cagaaaacat aaagattgtc ctccacctgt ccctgtgctc gcaattgtgc tcggagtcat
      301 tgcgggtatc gtaatcctcg gaattcttct cttgttgctc tggaaattgc tcacagtact
      361 tcatgataga tccgaggtac cggtagaaaa aatgagtaaa ggagaagaac ttttcactgg
      421 agttgtccca attcttgttg aattagatgg tgatgttaat gggcacaaat tttctgtcag
      481 tggagagggt gaaggtgatg caacatacgg aaaacttacc cttaaattta tttgcactac
      541 tggaaaacta cctgttccat gggtaagttt aaacatatat atactaacta accctgatta
      601 tttaaatttt cagccaacac ttgtcactac tttctgttat ggtgttcaat gcttctcgag
      661 atacccagat catatgaaac ggcatgactt tttcaagagt gccatgcccg aaggttatgt
      721 acaggaaaga actatatttt tcaaagatga cgggaactac aagacacgta agtttaaaca
      781 gttcggtact aactaaccat acatatttaa attttcaggt gctgaagtca agtttgaagg
      841 tgataccctt gttaatagaa tcgagttaaa aggtattgat tttaaagaag atggaaacat
      901 tcttggacac aaattggaat acaactataa ctcacacaat gtatacatca tggcagacaa
      961 acaaaagaat ggaatcaaag ttgtaagttt aaacatgatt ttactaacta actaatctga
     1021 tttaaatttt cagaacttca aaattagaca caacattgaa gatggaagcg ttcaactagc
     1081 agaccattat caacaaaata ctccaattgg cgatggccct gtccttttac cagacaacca
     1141 ttacctgtcc acacaatctg ccctttcgaa agatcccaac gaaaagagag accacatggt
     1201 ccttcttgag tttgtaacag ctgctgggat tacacatggc atggatgaac tatacaaata
     1261 gcattcgtag aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa
     1321 aaataatagg ggccgctgtc atcagagtaa gtttaaactg agttctacta actaacgagt
     1381 aatatttaaa ttttcagcat ctcgcgcccg tgcctctgac ttctaagtcc aattactctt
     1441 caacatccct acatgctctt tctccctgtg ctcccacccc ctatttttgt tattatcaaa
     1501 aaacttctct taatttcttt gttttttagc ttcttttaag tcacctctaa caatgaaatt
     1561 gtgtagattc aaaaatagaa ttaattcgta ataaaaagtc gaaaaaaatt gtgctccctc
     1621 cccccattaa taataattct atcccaaaat ctacacaatg ttctgtgtac acttcttatg
     1681 ttttttactt ctgataaatt tttttgaaac atcatagaaa aaaccgcaca caaaatacct
     1741 tatcatatgt tacgtttcag tttatgaccg caatttttat ttcttcgcac gtctgggcct
     1801 ctcatgacgt caaatcatgc tcatcgtgaa aaagttttgg agtatttttg gaatttttca
     1861 atcaagtgaa agtttatgaa attaattttc ctgcttttgc tttttggggt ttcccctatt
     1921 gtttgtcaag atttcgagga cggcgttttt cttgctaaaa tcacaagtat tgatgagcac
     1981 gatgcaagaa agatcggaag aaggtttggg tttgaggctc agtggaaggt gagtagaagt
     2041 tgataatttg aaagtggagt agtgtctatg gggtttttgc cttaaatgac agaatacatt
     2101 cccaatatac caaacataac tgtttcctac tagtcggccg tacgggccct ttcgtctcgc
     2161 gcgtttcggt gatgacggtg aaaacctctg acacatgcag ctcccggaga cggtcacagc
     2221 ttgtctgtaa gcggatgccg ggagcagaca agcccgtcag ggcgcgtcag cgggtgttgg
     2281 cgggtgtcgg ggctggctta actatgcggc atcagagcag attgtactga gagtgcacca
     2341 tatgcggtgt gaaataccgc acagatgcgt aaggagaaaa taccgcatca ggcggcctta
     2401 agggcctcgt gatacgccta tttttatagg ttaatgtcat gataataatg gtttcttaga
     2461 cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc tatttgttta tttttctaaa
     2521 tacattcaaa tatgtatccg ctcatgagac aataaccctg ataaatgctt caataatatt
     2581 gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc ccttattccc ttttttgcgg
     2641 cattttgcct tcctgttttt gctcacccag aaacgctggt gaaagtaaaa gatgctgaag
     2701 atcagttggg tgcacgagtg ggttacatcg aactggatct caacagcggt aagatccttg
     2761 agagttttcg ccccgaagaa cgttttccaa tgatgagcac ttttaaagtt ctgctatgtg
     2821 gcgcggtatt atcccgtatt gacgccgggc aagagcaact cggtcgccgc atacactatt
     2881 ctcagaatga cttggttgag tactcaccag tcacagaaaa gcatcttacg gatggcatga
     2941 cagtaagaga attatgcagt gctgccataa ccatgagtga taacactgcg gccaacttac
     3001 ttctgacaac gatcggagga ccgaaggagc taaccgcttt tttgcacaac atgggggatc
     3061 atgtaactcg ccttgatcgt tgggaaccgg agctgaatga agccatacca aacgacgagc
     3121 gtgacaccac gatgcctgta gcaatggcaa caacgttgcg caaactatta actggcgaac
     3181 tacttactct agcttcccgg caacaattaa tagactggat ggaggcggat aaagttgcag
     3241 gaccacttct gcgctcggcc cttccggctg gctggtttat tgctgataaa tctggagccg
     3301 gtgagcgtgg gtctcgcggt atcattgcag cactggggcc agatggtaag ccctcccgta
     3361 tcgtagttat ctacacgacg gggagtcagg caactatgga tgaacgaaat agacagatcg
     3421 ctgagatagg tgcctcactg attaagcatt ggtaactgtc agaccaagtt tactcatata
     3481 tactttagat tgatttaaaa cttcattttt aatttaaaag gatctaggtg aagatccttt
     3541 ttgataatct catgaccaaa atcccttaac gtgagttttc gttccactga gcgtcagacc
     3601 ccgtagaaaa gatcaaagga tcttcttgag atcctttttt tctgcgcgta atctgctgct
     3661 tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt gccggatcaa gagctaccaa
     3721 ctctttttcc gaaggtaact ggcttcagca gagcgcagat accaaatact gtccttctag
     3781 tgtagccgta gttaggccac cacttcaaga actctgtagc accgcctaca tacctcgctc
     3841 tgctaatcct gttaccagtg gctgctgcca gtggcgataa gtcgtgtctt accgggttgg
     3901 actcaagacg atagttaccg gataaggcgc agcggtcggg ctgaacgggg ggttcgtgca
     3961 cacagcccag cttggagcga acgacctaca ccgaactgag atacctacag cgtgagcatt
     4021 gagaaagcgc cacgcttccc gaagggagaa aggcggacag gtatccggta agcggcaggg
     4081 tcggaacagg agagcgcacg agggagcttc cagggggaaa cgcctggtat ctttatagtc
     4141 ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt gtgatgctcg tcaggggggc
     4201 ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg gttcctggcc ttttgctggc
     4261 cttttgctca catgttcttt cctgcgttat cccctgattc tgtggataac cgtattaccg
     4321 cctttgagtg agctgatacc gctcgccgca gccgaacgac cgagcgcagc gagtcagtga
     4381 gcgaggaagc ggaagagcgc ccaatacgca aaccgcctct ccccgcgcgt tggccgattc
     4441 attaatgcag ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa
     4501 ttaatgtgag ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc
     4561 gtatgttgtg tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg
     4621 attacgccaa gctgtaagtt taaacatgat cttactaact aactattctc atttaaattt
     4681 tcagagctta aaaatggctg aaatcactca caacgatgga tacgctaaca acttggaaat
     4741 gaaat
//
