LOCUS       sequence-276-exp        7172 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 7172)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 7172)
  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: L4664
FEATURES             Location/Qualifiers
     source          1..7172
                     /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(4612..4634)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4734..4753
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4831..4849)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4917..5021
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5022..5090,5091..5882)
                     /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(5240..5259)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6053..6641
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6542..6561
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6795..6812
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6929..6950
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6965..6970,6971..6988,6989..6995)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7003..7019
                     /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     7008..7030
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7027..7043
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7027..7043
                     /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 actttctcat ggggcgttca 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 atttcacaca acgtatacat cactgccgac aaacaaaaga atgggatcaa agcggtaagt
     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 gcatggatga actatacaaa tagcattcgt agaattgtaa gtttaaactt
     3661 tctcatacta actaacgcgc tctatttaaa ttttcagaat tccaactgag cgccggtcgc
     3721 taccattacc aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt
     3781 taaactgagt tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc
     3841 ctctgacttc taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc
     3901 ccacccccta tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc
     3961 ttttaagtca cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata
     4021 aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta
     4081 cacaatgttc tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc
     4141 atagaaaaaa ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa
     4201 tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa
     4261 gttttggagt atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg
     4321 cttttgcttt ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt
     4381 gctaaaatca caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt
     4441 gaggctcagt ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg
     4501 tttttgcctt aaatgacaga atacattccc aatataccaa acataactgt ttcctactag
     4561 tcggccgtac gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca
     4621 catgcagctc ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc
     4681 ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc
     4741 agagcagatt gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag
     4801 gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta
     4861 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg
     4921 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     4981 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     5041 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     5101 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     5161 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     5221 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     5281 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     5341 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     5401 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     5461 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     5521 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     5581 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     5641 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     5701 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     5761 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     5821 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     5881 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     5941 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     6001 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     6061 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     6121 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     6181 cgcagatacc aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact
     6241 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     6301 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     6361 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     6421 aactgagata cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg
     6481 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     6541 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     6601 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     6661 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     6721 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     6781 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     6841 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     6901 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     6961 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     7021 ttcacacagg aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt
     7081 actaactaac tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa
     7141 cgatggatac gctaacaact tggaaatgaa at
//
