LOCUS       sequence-282-exp       10279 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 10279)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 10279)
  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: L4683
FEATURES             Location/Qualifiers
     source          1..10279
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2511..2539)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3287..3307
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3440..3462)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7853..7875)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7975..7994
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(8072..8090)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8158..8262
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8263..8331,8332..9123)
                     /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(8481..8500)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9294..9882
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9783..9802
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     10036..10053
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    10170..10191
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(10206..10211,10212..10229,10230..10236)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10244..10260
                     /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     10249..10271
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(10268..10279,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(10268..10279,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa
       61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag
      121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc
      181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca
      241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat
      301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac
      361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt
      421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca
      481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc
      541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc
      601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta
      661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa
      721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg
      781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt
      841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt
      901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag
      961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa
     1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga
     1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca
     1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc
     1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta
     1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat
     1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg
     1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg
     1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca
     1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct
     1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat
     1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat
     1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa
     1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc
     1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa
     1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg
     1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc
     1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga
     2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt
     2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct
     2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc
     2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt
     2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa
     2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg
     2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gaggaccctt
     2461 ggagggtacc ggtagaaaaa atgagtaaag gagaagaact tttcactgga gttgtcccaa
     2521 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg
     2581 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac
     2641 ctgttccatg ggtaagttta aacatatata tactaactaa ccctgattat ttaaattttc
     2701 agccaacact tgtcactact ttcggttatg gtcttcaatg cttcgccaga tacccagatc
     2761 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     2821 ctatattttt caaagatgac gggaactaca agacacgtaa gtttaaacag ttcggtacta
     2881 actaaccata catatttaaa ttttcaggtg ctgaagtcaa gtttgaaggt gatacccttg
     2941 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     3001 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     3061 gaatcaaagt tgtaagttta aacatgattt tactaactaa ctaatctgat ttaaattttc
     3121 agaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     3181 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcct
     3241 accaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     3301 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaatgc ccggtgggtg
     3361 aagaccagaa acagcacctc gaactgagcc gcgatattgc ccagcgtttc aacgcgctgt
     3421 atggcgagat cgatcccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc
     3481 aacttaatcg ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc
     3541 gcaccgatcg cccttcccaa cagttgcgaa ggtaagttta aacagatcca tactaactaa
     3601 cttgttctga cataattttc agcttgaatg gcgaatggcg ctttgcctgg tttccggcac
     3661 cagaagcggt gccggaaagc tggctggagt gcgatcttcc tgaggccgat actgtcgtcg
     3721 tcccctcaaa ctggcagatg cacggttacg atgcgcccat ctacaccaac gtgacctatc
     3781 ccattacggt caatccgccg tttgttccca cggagaatcc gacgggttgt tactcgctca
     3841 catttaatgt tgatgaaagc tggctacagg aaggccagac gcgaattatt tttgatggcg
     3901 ttaactcggc gtttcatctg tggtgcaacg ggcgctgggt cggttacggc caggtaagtt
     3961 taattaagtt gatactaact aacaagatct gattaatttt caggacagtc gtttgccgtc
     4021 tgaatttgac ctgagcgcat ttttacgcgc cggagaaaac cgcctcgcgg tgatggtgct
     4081 gcgctggagt gacggcagtt atctggaaga tcaggatatg tggcggatga gcggcatttt
     4141 ccgtgacgtc tcgttgctgc ataaaccgac tacacaaatc agcgatttcc atgttgccac
     4201 tcgctttaat gatgatttct cccgcgctgt actggaggct gaagtccagg taagtttaaa
     4261 caggatctta ctaactaaca tgctaacact gaattttcag atgtgcggcg agttgcgtga
     4321 ctacctacgg gtaacagttt ctttatggca gggtgaaacg caggtcgcca gcggcaccgc
     4381 gcctttcggc ggtgaaatta tcgatgagcg tggtggttat gccgatcgcg tcacactacg
     4441 tctgaacgtc gaaaacccga aactgtggag cgccgaaatc ccgaatctct atcgtgcggt
     4501 ggttgaactg cacaccgccg acggcacgct gattgaagca gaagcctgcg atgtcggttt
     4561 ccgcgaggtg cggattgaaa atggtctgct gctgctgaac ggcaagccgt tgctgattcg
     4621 aggcgttaac cgtcacgagc atcatcctct gcatggtcag gtcatggatg agcagacgat
     4681 ggtgcaggat gtaagtttaa actattcgtt actaactaac tttaaacatt taaattttca
     4741 gatcctgctg atgaagcaga acaactttaa cgccgtgcgc tgttcgcatt atccgaacca
     4801 tccgctgtgg tacacgctgt gcgaccgcta cggcctgtat gtggtggatg aagccaatat
     4861 tgaaacccac ggcatggtgc caatgaatcg tctgaccgat gatccgcgct ggctaccggc
     4921 gatgagcgaa cgcgtaacgc gaatggtgca gcgcgatcgt aatcacccga gtgtgatcat
     4981 ctggtcgctg gggaatggta agtttaaaca gttgaatact aactaacgga gatctttgaa
     5041 attttcagaa tcaggccacg gcgctaatca cgacgcgctg tatcgctgga tcaaatctgt
     5101 cgatccttcc cgcccggtgc agtatgaagg cggcggagcc gacaccacgg ccaccgatat
     5161 tatttgcccg atgtacgcgc gcgtggatga agaccagccc ttcccggctg tgccgaaatg
     5221 gtccatcaaa aaatggcttt cgctacctgg agagacgcgc ccgctgattc tttgcgaggt
     5281 aagtttaaac agaactctac taactaacac attagatcct aattttcagt acgctcacgc
     5341 gatgggcaac agtcttggcg gtttcgctaa atactggcag gcgtttcgtc agtatccccg
     5401 tttacagggc ggcttcgtct gggactgggt ggatcagtcg ctgattaaat atgatgaaaa
     5461 cggcaacccg tggtcggctt acggcggtga ttttggcgat acgccgaacg atcgccagtt
     5521 ctgtatgaac ggtctggtct ttgccgaccg cacgccgcat ccagcgtaag tttaaacaat
     5581 aacctactaa ctaacgtaga taatttaaat tttcaggctg acggaagcaa aacaccagca
     5641 gcagtttttc cagttccgtt tatccgggca aaccatcgaa gtgaccagcg aatacctgtt
     5701 ccgtcatagc gataacgagc tcctgcactg gatggtggcg ctggatggta agccgctggc
     5761 aagcggtgaa gtgcctctgg atgtcgctcc acaaggtaaa cagttgattg aactgcctga
     5821 actaccgcag ccggagagcg ccgggcaact ctggctcaca gtacgcgtag tgcaaccgaa
     5881 cgcgaccgca tggtcagaag ccgggcacat cagcgcatgg cagcagtgga ggtaagttta
     5941 aacaagatcc tactaactaa ctctacattg atgaattttc agactggcgg aaaacctcag
     6001 tgtgacgctc cccgccgcgt cccacgccat cccgcatctg accaccagcg aaatggattt
     6061 ttgcatcgag ctgggtaata agcgttggca atttaaccgc cagtcaggct ttctttcaca
     6121 gatgtggatt ggcgataaaa aacaactgct gacgccgctg cgcgatcagt tcacccgtgc
     6181 accgctggat aacgacattg gcgtaagtga agcgacccgc attgacccta acgcctgggt
     6241 cggtaagttt aaacaaagtt gtactaacta acgaagatct tgataatttt cagaacgctg
     6301 gaaggcggcg ggccattacc aggccgaagc agcgttgttg cagtgcacgg cagatacact
     6361 tgctgatgcg gtgctgatta cgaccgctca cgcgtggcag catcagggga aaaccttatt
     6421 tatcagccgg aaaacctacc ggattgatgg tagtggtcaa atggcgatta ccgttgatgt
     6481 tgaagtggcg agcgatacac cgcatccggc gcggattggc ctgaactgcc agctggcgca
     6541 ggtagcagag cgggtaaact ggctcggatt agggccgcaa gaaaactatc ccgaccgcct
     6601 tactgccgcc tgttttgacc gctgggatct gccattgtca gacatgtagt aagtttaaac
     6661 ttgatagtac taactaacat gtttcattta aattttcagt accccgtacg tcttcccgag
     6721 cgaaaacggt ctgcgctgcg ggacgcgcga attgaattat ggcccacacc agtggcgcgg
     6781 cgacttccag ttcaacatca gccgctacag tcaacagcaa ctgatggaaa ccagccatcg
     6841 ccatctgctg cacgcggaag aaggcacatg gctgaatatc gacggtttcc atatggggat
     6901 tggtggcgac gactcctgga gcccgtcagt atcggcggaa ttccaactga gcgccggtcg
     6961 ctaccattac caacttgtct ggtgtcaaaa ataatagggg ccgctgtcat cagagtaagt
     7021 ttaaactgag ttctactaac taacgagtaa tatttaaatt ttcagcatct cgcgcccgtg
     7081 cctctgactt ctaagtccaa ttactcttca acatccctac atgctctttc tccctgtgct
     7141 cccaccccct atttttgtta ttatcaaaaa acttctctta atttctttgt tttttagctt
     7201 cttttaagtc acctctaaca atgaaattgt gtagattcaa aaatagaatt aattcgtaat
     7261 aaaaagtcga aaaaaattgt gctccctccc cccattaata ataattctat cccaaaatct
     7321 acacaatgtt ctgtgtacac ttcttatgtt ttttacttct gataaatttt tttgaaacat
     7381 catagaaaaa accgcacaca aaatacctta tcatatgtta cgtttcagtt tatgaccgca
     7441 atttttattt cttcgcacgt ctgggcctct catgacgtca aatcatgctc atcgtgaaaa
     7501 agttttggag tatttttgga atttttcaat caagtgaaag tttatgaaat taattttcct
     7561 gcttttgctt tttggggttt cccctattgt ttgtcaagat ttcgaggacg gcgtttttct
     7621 tgctaaaatc acaagtattg atgagcacga tgcaagaaag atcggaagaa ggtttgggtt
     7681 tgaggctcag tggaaggtga gtagaagttg ataatttgaa agtggagtag tgtctatggg
     7741 gtttttgcct taaatgacag aatacattcc caatatacca aacataactg tttcctacta
     7801 gtcggccgta cgggcccttt cgtctcgcgc gtttcggtga tgacggtgaa aacctctgac
     7861 acatgcagct cccggagacg gtcacagctt gtctgtaagc ggatgccggg agcagacaag
     7921 cccgtcaggg cgcgtcagcg ggtgttggcg ggtgtcgggg ctggcttaac tatgcggcat
     7981 cagagcagat tgtactgaga gtgcaccata tgcggtgtga aataccgcac agatgcgtaa
     8041 ggagaaaata ccgcatcagg cggccttaag ggcctcgtga tacgcctatt tttataggtt
     8101 aatgtcatga taataatggt ttcttagacg tcaggtggca cttttcgggg aaatgtgcgc
     8161 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     8221 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     8281 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     8341 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     8401 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     8461 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     8521 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     8581 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     8641 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     8701 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     8761 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     8821 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     8881 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     8941 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     9001 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     9061 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     9121 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     9181 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     9241 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     9301 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     9361 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     9421 gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca cttcaagaac
     9481 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     9541 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     9601 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     9661 gaactgagat acctacagcg tgagcattga gaaagcgcca cgcttcccga agggagaaag
     9721 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     9781 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     9841 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     9901 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     9961 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
    10021 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
    10081 ccgcctctcc ccgcgcgttg gccgattcat taatgcagct ggcacgacag gtttcccgac
    10141 tggaaagcgg gcagtgagcg caacgcaatt aatgtgagtt agctcactca ttaggcaccc
    10201 caggctttac actttatgct tccggctcgt atgttgtgtg gaattgtgag cggataacaa
    10261 tttcacacag gaaacagct
//
