LOCUS       sequence-120-exp       10549 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 10549)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 10549)
  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: BK48
FEATURES             Location/Qualifiers
     source          1..10549
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1207..1235)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1983..2003
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(7734..7753)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(7735..7753)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(7760..7777)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7760..7776)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7769..7791)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      7918..8373
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(8005..8024)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     8215..8236
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        8400..8504
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(8505..8573,8574..9365)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAISMSDNTAANLLLTTIGGPKELTAFFHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8723..8742)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9536..10124
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     10278..10295
                     /label=L4440
                     /note="L4440 vector, forward primer"
     promoter        join(10448..10453,10454..10471,10472..10478)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    10486..10502
                     /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     10491..10513
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     10510..10526
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     10510..10526
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     promoter        join(10547..10549,1..16)
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 taaccctcac taaagggaac aaaagctggc tccaaaatcg tcttccgctc tgaaaaacga
       61 aagtggacct ttgacatccg aaaaaatggg cgaaaaaatg aaattgagct ttttgggtcg
      121 aaaaaaatgt ttttagaatg ctgagaacac gttaaacacg aagatcatat ttattttgag
      181 acccggatgc tctgaaaatg tctgacatag atttaaaaaa gcatatatat atttttcatt
      241 ttcaacgtga aagttttgtg caactttata gaatctccta ttggcacatt gttttttatt
      301 taactgaggc agtttttgaa cacctttttg aaactttgaa tctctttgaa gtatactgtc
      361 gaaaagactg acttgagcgt tcgaaatgcc agaagaaaac tatatttgaa tctcgcgcta
      421 aattgagaaa tgcaaccgcg ctccactgga caattggaaa aaaaatttat tcggaggcga
      481 caacggtatt ttcgaaattg attttctgtg tattttctca ttttttataa attcttcttt
      541 gatttatcgt tcgtttgtga gaaatttaat tgtattcaaa cttttttata gtaagatgtc
      601 gtcgagtcgg tcacaatcac ctgaaactcc aaaggcagcc agtgaggaac gtgaagaaga
      661 agaaaaagag tcatctgaac aggtttgatt ttctttctgg tcaaaaagat gaaattattg
      721 attttcagcc agatactccc aaaactagca gcgagaagtc tgcaagtcgt tcacagtcgc
      781 ccagagaatc gcgggaagtg agccaagagg tatgtttttc aaaaatcaat aactgatcat
      841 aatttttatt gtttggtgaa tttaagaaaa taatattcga aaattcctct gaattatcaa
      901 gattgcagta ttaatttcga gaaaaattga gatattcata gagctattgt aaattttctt
      961 gatttcagac tgaaacttcg gaaaatcaag agaaaatcaa agaaaaggat gacggggatg
     1021 atcagcctgg cacaccgaac agctatagaa gccgggaaac ttcaccagct ccaaaaaggt
     1081 ccaaggagac caggtttgtc aaaagcttcc tgcgattaat tctcatttca atttttcaga
     1141 gaatcagagt ctcctgaaaa atcaccggta gaaaaaatga gtaaaggaga agaacttttc
     1201 actggagttg tcccaattct tgttgaatta gatggtgatg ttaatgggca caaattttct
     1261 gtcagtggag agggtgaagg tgatgcaaca tacggaaaac ttacccttaa atttatttgc
     1321 actactggaa aactacctgt tccatgggta agtttaaaca tatatatact aactaaccct
     1381 gattatttaa attttcagcc aacacttgtc actactttct gttatggtgt tcaatgcttc
     1441 tcgagatacc cagatcatat gaaacggcat gactttttca agagtgccat gcccgaaggt
     1501 tatgtacagg aaagaactat atttttcaaa gatgacggga actacaagac acgtaagttt
     1561 aaacagttcg gtactaacta accatacata tttaaatttt caggtgctga agtcaagttt
     1621 gaaggtgata cccttgttaa tagaatcgag ttaaaaggta ttgattttaa agaagatgga
     1681 aacattcttg gacacaaatt ggaatacaac tataactcac acaatgtata catcatggca
     1741 gacaaacaaa agaatggaat caaagttgta agtttaaaca tgattttact aactaactaa
     1801 tctgatttaa attttcagaa cttcaaaatt agacacaaca ttgaagatgg aagcgttcaa
     1861 ctagcagacc attatcaaca aaatactcca attggcgatg gccctgtcct tttaccagac
     1921 aaccattacc tgtccacaca atctgccctt tcgaaagatc ccaacgaaaa gagagaccac
     1981 atggtccttc ttgagtttgt aacagctgct gggattacac atggcatgga tgaactatac
     2041 aaatgcccgg ttcgttcaag atctcccaga aggtcttcag cacgttcccc gtcacgatct
     2101 cctagacggc gccgagaaag aagctcagaa agaaagcaat ccgaagagcc agcaccgcta
     2161 ccagagaaaa agaagaaaga gccgctggat attctacgaa caagaaccgg aggagcatat
     2221 attccacccg ccaaacttcg acttatgcaa caacagatta gtgataagca aagtgaacag
     2281 tatcagagaa tgaattggga aagaatgaag aaaaagattc acggattggt taacagagtc
     2341 aacgcgaaga atcttgttca aattgtcaga gaacttcttc aagagaatgt gattcgttca
     2401 aagtgagtga gaaaatcgaa ggaaaaggaa agaattaatt taatttttca ggggacttct
     2461 ctgccgtgac attattcaag ctcaggcttt ctcaccagga ttctctaacg tctatgcagc
     2521 tttggcggca gttatcaact cgaaattccc tcatgtcggt gaacttcttc tccgtcgtct
     2581 gattgtacag ttcaaaagaa gtttccgtag aaatgacaga ggcgtcacgg tgaacgtgat
     2641 caaattcatc gcacatttga ttaatcaaca agttgctcac gaagttcttg cgctggaaat
     2701 catgattctg atgcttgaag aaccaactga tgattcagtt gaagtcgcca ttgcgttcct
     2761 gaaagagtgt ggagcaaagc ttctggagat tgctccagca gctcttaaca gtgtctacga
     2821 ccgtcttcgt gcaattctca tggaaactga aagatcggaa aatgcactgg atcgacgtat
     2881 tcagtatatg attgagactg caatgcagat tcgaaaggac aaatttgcgg taaggtagaa
     2941 tatataaata gtttattaga aaaaaataaa ttagaataat ttaaattcct actagccaat
     3001 caggcgacct ttttgcgcat agttctatta ttgaaaaatt tggagaattt ctcatattct
     3061 cgctcggaaa tctggaattc gacgagatct tctggcttct gtgcagctgc atcgctttgt
     3121 gctccctttc tcgcttgtct tctgtgtaca ccaagaacct tgttgagttc atcaactgaa
     3181 tctgtgactg gcttgttgct cactggatgc actagacgac tgattctcga gaaatcagat
     3241 tgagttgcga ttagggtgac ctagaaattg ggaataatac gaacttttga aaatattcag
     3301 gaggattaaa aaaattattc tcgacaatcc tacaaattta cttattgcac catgttgctc
     3361 caacattttt cattaaaagt taatgaaaaa atgtagaaaa tcggaaattg gcaattttca
     3421 gaccattttt aagcattttc aaaaaaaaat tgcagctgaa ataaatgtca ttttcagata
     3481 aatcgagcga ttttctgttg tctgacacta gtttttagtt ttaaaaaatg ttggaagaac
     3541 atggtgcaat aggtaatttc atagaatttc catgtgtttt ttttcaatta accaattatc
     3601 caaatcttcc aaactcacat tttgcggagc tgggctatca agaatctgct gcagttttat
     3661 aagacgagca tctctgatat cactgaaaat taatttttaa tcaaaacttg aatatcaact
     3721 aaacccactt attaactttc tcgatcttct gtcgttcggt acgatgacgg tgaagaagcc
     3781 aattgtagta gttgatttgg ttcaagtcct ttcggtgttg tacgtcagtg tcctgcaatg
     3841 ctatttagtt ataacttagg cctaagattc aatttaatga agtgattaaa tttgttctct
     3901 gaacctctta agatgatctt ttggattaga aacatataag acaggtttac ctatctatta
     3961 aaaaacagat caaaatagat acgaccaaat cggataatcc atgcctacct ggcatctagg
     4021 aacgtgttct tagaagattt cttacgtaat cgtatgaaga aataacaatt tgatcgttgg
     4081 ccagcaaaaa tagggtttta agtgggatag tgtttttatt agctaaccgg aaaattttat
     4141 agtttttttt tgcaagaaac cactgaaaac cccctaattg tatacatttt ttggagcagc
     4201 ttctggtctt tttgagcaat aaaattcgat aaaacagaat ttaagtgtaa attgttcaca
     4261 tttagtttct attttatcaa attttgttgc tcaaaaacat tcgaagctgc tctaaaaaaa
     4321 tgcattaaaa aaggggtttt cagtggtttt tcacattaaa aaagctaatt ttaactaaaa
     4381 atccatcata tttccaactt tgtcacaaca ataaaatgct ggtcaaaatg tgttcgaaaa
     4441 aatgtttttt tttttaattt ttataattta aaaatagttt tctttcgctg ggacacatac
     4501 atttttgggc gtaaattttc agttcaaatt tccattttta caaccataat cataaagcta
     4561 cgtctgatct ctctcgcact tacctgcgcc tgattcgaaa gaacaaccgt agccaaaaga
     4621 acaagaagaa caagcacgta gttgtggtag tggacgttca tcacgcaata ctgaccaatg
     4681 gtcgtggggt ctcactttcc gtactattga gagaggggag actgaagatg gcaattgagg
     4741 acagtgtctt cgacgcacgc atgcatccat aagcataatc caggagggat ggagagaaaa
     4801 atcttgtttc taagcccctc cctttgtaat acatacacat atctaatacc gaagaatggc
     4861 taattgaatg gacgtcagct gttgctgtag ttgccaaggc atcatcgatg aaataactga
     4921 aagaaagaat taaataatta ttgcaggcgt atccggcggt cattgaagac ttggacttga
     4981 ttgaggagga ggatcagatc atccatacac ttaatttgga ggatgcggtt gatccggaaa
     5041 atgggcttag taagtgactg accacacgcg gggggcatta atttaataaa ttgaattcca
     5101 tttcagatgt gttcaaacta gatccagaat tcgaaaagaa cgaggaggtt tatgaggaga
     5161 tccgtaagga aatcattgga aacgccgata tttcggatga ggatggtggc gacgagttgg
     5221 atgatgaaga agagggtagt gatgtggaag aggctccgaa gaagactaca gagattattg
     5281 ataatactga tcagaattga ctgctttcag aaggtattca ttttgagttt tgggccggca
     5341 aatctgtaag ttgccggttg ccgaaaattt gctgaatttg ccggaaaaaa aaattccgga
     5401 atttatttaa aaactttttg taaaaattaa attaaatttg caacttttca gagaagtcta
     5461 cctgacaatg caatcatctt tggactacca agaagctgct cacaaattgc tgaaaatgaa
     5521 gattccagac agcatgcagg tcagcgatgt tgcaaagaaa aattttcgac caaaaaaacc
     5581 aaccaatcat aaaatttaaa aaaaaactcc gtttttttct ttttttttat acgagaaaaa
     5641 ccaaaaaaat gtatttttgc caaattctaa aatactatcc ccgaaatttt caatattttc
     5701 tctttcagaa cgaactctgc gcgatgcttg tcgattgttg tgctcaacag cgtacctacg
     5761 agcgattcta cggaatgctc atcgaacgtt tctgccgact tcgcctcgaa taccagcaat
     5821 actttgaaaa gctctgccag gacacgtatt ccacgattca ccgaattgac atcacaaaac
     5881 tgcggaattt ggctcgcctt attgctcatt tgctctcgac ggatgctatt gactggaaga
     5941 ttttggccga tatgaaaatg accgaagagg acacaacttc ttctggcaga atctatatta
     6001 aatatatatt taatgaactt gtggaggcga tgggaatggt taaacttcat tcgagagtta
     6061 ctgatccgtg agtttcctag agagagttgt tttcgtattc aattttccct attttcagaa
     6121 ctttggctca ttgctttgtt ggattattcc cacgaactaa tccgaacagc gcacgatttt
     6181 cgatcaactt cttcacaatg attggattgg gtggtttgac gttggaactt cgtgaatggc
     6241 tggcaaaggg tctcaagaag aagaagggaa tgctggatca gttgaaggcc gaatcaagct
     6301 cagattcatc gtcgtcttcg gattcgtcag actcgtctga ttcttcggat tctgacgatt
     6361 catccgactc gtcttcagat tcctcatctt cttcagaatc agagccagaa ccaccgaaga
     6421 aaaagaagaa gaagaacagt gaagagagtt ccaaaaagaa ggaaaaagag aatattggtc
     6481 gacgggatcg tggagacaag agagctgaac gtcatcgtga tcaaagtgtg gagaacaagg
     6541 acaaggatcg tcgacgtcgc caggattctg acgaaaatcg tcggccagaa cgaggagatg
     6601 accgcaagga tcggagtaaa gatcgtcgtc gtcaagactc ggatgatgag gatcggaaag
     6661 gtcgtgaacg tcgggaagat tcaggggaaa gacgtcgcgg agatcgggat cgacgtgatc
     6721 gaaacaagga tcaggaggat caccgtgaag atcgccgtga ccgaagcaag gatcgtgagg
     6781 atcgacgtga tcgccgtcgt catgactctg atgatgatcg taaaactcgt cgggatagaa
     6841 gtgaagagcg aggaggacgt cgtcgtgaag tggaatcgga tgatcgacgc cgacgtcgtt
     6901 gaattttcaa attttaaata ctgaatattt gttttttttc ctattattta tttattctct
     6961 ttgtgttttt tttcttgctt tctaaaaaat taattcaatc caaatctaaa catttttttt
     7021 tctctttccg tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt
     7081 atttatcttc tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca
     7141 cacaaaagga tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga
     7201 aatatggagg aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt
     7261 aaaaaataag caaagaaatt aggcgaacga tgagaattgt cctcgcttgg caaatgcgaa
     7321 tccgtatgga gaggcacgtt tggcgaaggc aaatgttcgg tatggagatc tgtaaaaatt
     7381 tttaagttga aatttggtgt tgctctttta caaaattttc cgattttcgc ttgaaattac
     7441 ggtgccaggt ctcgacacgt cttccaattt ttcaaattca aaagagcctt taatgggctg
     7501 tagttgctaa tttctcgttt ttgaaaattt ttcttccgtt taatcgaaat ttgatgtatt
     7561 ttatttatga ttttcaataa atttcaaaga aactggtgaa aactcggaaa attgtgaact
     7621 acagtaatcc aatccttaaa ggcgcacacc ttttaaatgt ccgccccaat acgatatttt
     7681 tttaagattc gggccctcta gagcggccgc caccgcggtg gagctccaat tcgccctata
     7741 gtgagtcgta ttacaattca ctggccgtcg ttttacaacg tcgtgactgg gaaaaccctg
     7801 gcgttaccca acttaatcgc cttgcagcac atcccccctt cgccagctgg cgtaatagcg
     7861 aagaggcccg caccgatcgc ccttcccaac agttgcgtag cctgaatggc gaatgggacg
     7921 cgccctgtag cggcgcatta agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta
     7981 cacttgccag cgccctagcg cccgctcctt tcgctttctt cccttccttt ctcgccacgt
     8041 tcgccggctt tccccgtcaa gctctaaatc gggggctccc tttagggttc cgatttagtg
     8101 ctttacggca cctcgacccc aaaaaacttg attagggtga tggttcacgt agtgggccat
     8161 cgccctgata gacggttttt cgccctttga cgttggagtc cacgttcttt aatagtggac
     8221 tcttgttcca aactggaaca acactcaacc ctatctcggt ctattctttt gatttataag
     8281 ggattttgcc gatttcggcc tattggttaa aaaatgagct gatttaacaa aaatttaacg
     8341 cgaattttaa caaaatatta acgtttacaa tttcaggtgg cacttttcgg ggaaatgtgc
     8401 gcggaacccc tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac
     8461 aataaccctg ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt
     8521 tccgtgtcgc ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag
     8581 aaacgctggt gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg
     8641 aactggatct caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa
     8701 tgatgagcac ttttaaagtt ctgctatgtg gcgcggtatt atcccgtatt gacgccgggc
     8761 aagagcaact cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag
     8821 tcacagaaaa gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa
     8881 gcatgagtga taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc
     8941 taaccgcttt ttttcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg
     9001 agctgaatga agccatacca aacgacgagc gtgacaccac gatgcctgta gcaatggcaa
     9061 caacgttgcg caaactatta actggcgaac tacttactct agcttcccgg caacaattaa
     9121 tagactggat ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg
     9181 gctggtttat tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag
     9241 cactggggcc agatggtaag ccctcccgta tcgtagttat ctacacgacg ggcagtcagg
     9301 caactatgga tgaacgaaat agacagatcg ctgagatagg tgcctcactg attaagcatt
     9361 ggtaactgtc agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt
     9421 aatttaaaag gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac
     9481 gtgagttttc gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag
     9541 atcctttttt tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg
     9601 tggtttgttt gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca
     9661 gagcgcagat accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga
     9721 actctgtagc accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca
     9781 gtggcgataa gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc
     9841 agcggtcggg ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca
     9901 ccgaactgag atacctacag cgtgagcatt gagaaagcgc cacgcttccc gaagggagaa
     9961 aggcggacag gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc
    10021 caggggggaa cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc
    10081 gtcgattttt gtgatgctcg tcaggggggc cgagcctatg gaaaaacgcc agcaacgcgg
    10141 cctttttacg gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat
    10201 cccctgattc tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca
    10261 gccgaacgac cgagcgcagc gagtcagtga gcgaggaagc ggaagagcgc ccaatacgca
    10321 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
    10381 actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag ttacctcact cattaggcac
    10441 cccaggcttt acactttatg cttccggctc ctatgttgtg tggaattgtg agcggataac
    10501 aatttcacac aggaaacagc tatgaccatg attacgccaa gctcggaat
//
