LOCUS       sequence-243-exp        6708 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 6708)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6708)
  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: L4057
FEATURES             Location/Qualifiers
     source          1..6708
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2794..2822)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3570..3590
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4148..4170)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4270..4289
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4367..4385)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4453..4557
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4558..4626,4627..5418)
                     /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(4776..4795)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5589..6177
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6078..6097
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6331..6348
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6465..6486
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6501..6506,6507..6524,6525..6531)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6539..6555
                     /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     6544..6566
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6563..6579
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6563..6579
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     2461 atagaacatt ttcaggagga cccttggagg gtaccgccgg caatgccacc ttcaacatca
     2521 ttgctgctcc tcgcagcact tcttccattc gctttaccag caagcgattg gaagactgga
     2581 gaagtcactc cggataatgc aacagtctgg gtcagaaaac ataaagattg tcctccacct
     2641 gtccctgtgc tcgcaattgt gctcggagtc attgcgggta tcgtaatcct cggaattctt
     2701 ctcttgttgc tctggaaatt gctcacagta cttcatgata gatccgaggt accggtagaa
     2761 aaaatgagta aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat
     2821 ggtgatgtta atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac
     2881 ggaaaactta cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt
     2941 ttaaacatat atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact
     3001 actttctgtt atggtgttca atgcttctcg agatacccag atcatatgaa acagcatgac
     3061 tttttcaaga gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat
     3121 gacgggaact acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt
     3181 aaattttcag gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta
     3241 aaaggtattg attttaaaga agatggaaac attcttggac acaaattgga atacaactat
     3301 aactcacaca atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt
     3361 ttaaacatga ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga
     3421 cacaacattg aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt
     3481 ggcgatggcc ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg
     3541 aaagatccca acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg
     3601 attacacatg gcatggacga actatacaaa agcccacaag ctttggaatt cgctagccgg
     3661 ccatacaagt aatccggatg atcgacgcca acgtcgttga attttcaaat tttaaatact
     3721 gaatatttgt tttttttcct attatttatt tattctcttt gtgttttttt tcttgctttc
     3781 taaaaaatta attcaatcca aatctaaaca tttttttttc tctttccgtc tcccaattcg
     3841 tattccgctc ctctcatctg aacacaatgt gcaagtttat ttatcttctc gctttcattt
     3901 cattaggacg tggggggaat tggtggaagg gggaaacaca caaaaggatg atggaaatga
     3961 aataaggaca cacaatatgc aacaacattc aattcagaaa tatggaggaa ggtttaaaag
     4021 aaaacataaa aatatataga ggaggaagga aaactagtaa aaaataagca aagaaattag
     4081 gcgaacgatg agaattgtcc tcgcttgggc cctttcgtct cgcgcgtttc ggtgatgacg
     4141 gtgaaaacct ctgacacatg cagctcccgg agacggtcac agcttgtctg taagcggatg
     4201 ccgggagcag acaagcccgt cagggcgcgt cagcgggtgt tggcgggtgt cggggctggc
     4261 ttaactatgc ggcatcagag cagattgtac tgagagtgca ccatatgcgg tgtgaaatac
     4321 cgcacagatg cgtaaggaga aaataccgca tcaggcggcc ttaagggcct cgtgatacgc
     4381 ctatttttat aggttaatgt catgataata atggtttctt agacgtcagg tggcactttt
     4441 cggggaaatg tgcgcggaac ccctatttgt ttatttttct aaatacattc aaatatgtat
     4501 ccgctcatga gacaataacc ctgataaatg cttcaataat attgaaaaag gaagagtatg
     4561 agtattcaac atttccgtgt cgcccttatt cccttttttg cggcattttg ccttcctgtt
     4621 tttgctcacc cagaaacgct ggtgaaagta aaagatgctg aagatcagtt gggtgcacga
     4681 gtgggttaca tcgaactgga tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa
     4741 gaacgttttc caatgatgag cacttttaaa gttctgctat gtggcgcggt attatcccgt
     4801 attgacgccg ggcaagagca actcggtcgc cgcatacact attctcagaa tgacttggtt
     4861 gagtactcac cagtcacaga aaagcatctt acggatggca tgacagtaag agaattatgc
     4921 agtgctgcca taaccatgag tgataacact gcggccaact tacttctgac aacgatcgga
     4981 ggaccgaagg agctaaccgc ttttttgcac aacatggggg atcatgtaac tcgccttgat
     5041 cgttgggaac cggagctgaa tgaagccata ccaaacgacg agcgtgacac cacgatgcct
     5101 gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg aactacttac tctagcttcc
     5161 cggcaacaat taatagactg gatggaggcg gataaagttg caggaccact tctgcgctcg
     5221 gcccttccgg ctggctggtt tattgctgat aaatctggag ccggtgagcg tgggtctcgc
     5281 ggtatcattg cagcactggg gccagatggt aagccctccc gtatcgtagt tatctacacg
     5341 acggggagtc aggcaactat ggatgaacga aatagacaga tcgctgagat aggtgcctca
     5401 ctgattaagc attggtaact gtcagaccaa gtttactcat atatacttta gattgattta
     5461 aaacttcatt tttaatttaa aaggatctag gtgaagatcc tttttgataa tctcatgacc
     5521 aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag accccgtaga aaagatcaaa
     5581 ggatcttctt gagatccttt ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca
     5641 ccgctaccag cggtggtttg tttgccggat caagagctac caactctttt tccgaaggta
     5701 actggcttca gcagagcgca gataccaaat actgtccttc tagtgtagcc gtagttaggc
     5761 caccacttca agaactctgt agcaccgcct acatacctcg ctctgctaat cctgttacca
     5821 gtggctgctg ccagtggcga taagtcgtgt cttaccgggt tggactcaag acgatagtta
     5881 ccggataagg cgcagcggtc gggctgaacg gggggttcgt gcacacagcc cagcttggag
     5941 cgaacgacct acaccgaact gagataccta cagcgtgagc attgagaaag cgccacgctt
     6001 cccgaaggga gaaaggcgga caggtatccg gtaagcggca gggtcggaac aggagagcgc
     6061 acgagggagc ttccaggggg aaacgcctgg tatctttata gtcctgtcgg gtttcgccac
     6121 ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac
     6181 gccagcaacg cggccttttt acggttcctg gccttttgct ggccttttgc tcacatgttc
     6241 tttcctgcgt tatcccctga ttctgtggat aaccgtatta ccgcctttga gtgagctgat
     6301 accgctcgcc gcagccgaac gaccgagcgc agcgagtcag tgagcgagga agcggaagag
     6361 cgcccaatac gcaaaccgcc tctccccgcg cgttggccga ttcattaatg cagctggcac
     6421 gacaggtttc ccgactggaa agcgggcagt gagcgcaacg caattaatgt gagttagctc
     6481 actcattagg caccccaggc tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt
     6541 gtgagcggat aacaatttca cacaggaaac agctatgacc atgattacgc caagctgtaa
     6601 gtttaaacat gatcttacta actaactatt ctcatttaaa ttttcagagc ttaaaaatgg
     6661 ctgaaatcac tcacaacgat ggatacgcta acaacttgga aatgaaat
//
