LOCUS       sequence-142-exp        8272 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 8272)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 8272)
  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: L3136
FEATURES             Location/Qualifiers
     source          1..8272
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             307..327
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(370..398)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     1146..1166
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(1299..1321)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5712..5734)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5834..5853
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5931..5949)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        6017..6121
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(6122..6190,6191..6982)
                     /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(6340..6359)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      7153..7741
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7642..7661
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7895..7912
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    8029..8050
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(8065..8070,8071..8088,8089..8095)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8103..8119
                     /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     8108..8130
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     8127..8143
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     8127..8143
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgac tctagacact tttcagctac ctagatacat
       61 ggatatcccc gcctcccaat ccacccaccc agggaaaaag aagggctcgc cgaaaatcaa
      121 agttatctcc aggctcgcgc atcccaccga gcggttgact tctctccacc acttttcatt
      181 ttaaccctcg gggtacggga ttggccaaag gacccaaagg tatgtttcga atgatactaa
      241 cataacatag aacattttca ggaggaccct tcttggaggg taccgagctc agaaaaaatg
      301 actgctccaa agaagaagcg taaggtaccg gtagaaaaaa tgagtaaagg agaagaactt
      361 ttcactggag ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt
      421 tctgtcagtg gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt
      481 tgcactactg gaaaactacc tgttccatgg gtaagtttaa acatatatat actaactaac
      541 cctgattatt taaattttca gccaacactt gtcactactt tctgttatgg tgttcaatgc
      601 ttctcgagat acccagatca tatgaaacgg catgactttt tcaagagtgc catgcccgaa
      661 ggttatgtac aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtaag
      721 tttaaacagt tcggtactaa ctaaccatac atatttaaat tttcaggtgc tgaagtcaag
      781 tttgaaggtg atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat
      841 ggaaacattc ttggacacaa attggaatac aactataact cacacaatgt atacatcatg
      901 gcagacaaac aaaagaatgg aatcaaagtt gtaagtttaa acatgatttt actaactaac
      961 taatctgatt taaattttca gaacttcaaa attagacaca acattgaaga tggaagcgtt
     1021 caactagcag accattatca acaaaatact ccaattggcg atggccctgt ccttttacca
     1081 gacaaccatt acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac
     1141 cacatggtcc ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta
     1201 tacaaatgcc cggtgggtga agaccagaaa cagcacctcg aactgagccg cgatattgcc
     1261 cagcgtttca acgcgctgta tggcgagatc gatcccgtcg ttttacaacg tcgtgactgg
     1321 gaaaaccctg gcgttaccca acttaatcgc cttgcagcac atcccccttt cgccagctgg
     1381 cgtaatagcg aagaggcccg caccgatcgc ccttcccaac agttgcgaag gtaagtttaa
     1441 acagatccat actaactaac ttgttctgac ataattttca gcttgaatgg cgaatggcgc
     1501 tttgcctggt ttccggcacc agaagcggtg ccggaaagct ggctggagtg cgatcttcct
     1561 gaggccgata ctgtcgtcgt cccctcaaac tggcagatgc acggttacga tgcgcccatc
     1621 tacaccaacg tgacctatcc cattacggtc aatccgccgt ttgttcccac ggagaatccg
     1681 acgggttgtt actcgctcac atttaatgtt gatgaaagct ggctacagga aggccagacg
     1741 cgaattattt ttgatggcgt taactcggcg tttcatctgt ggtgcaacgg gcgctgggtc
     1801 ggttacggcc aggtaagttt aattaagttg atactaacta acaagatctg attaattttc
     1861 aggacagtcg tttgccgtct gaatttgacc tgagcgcatt tttacgcgcc ggagaaaacc
     1921 gcctcgcggt gatggtgctg cgctggagtg acggcagtta tctggaagat caggatatgt
     1981 ggcggatgag cggcattttc cgtgacgtct cgttgctgca taaaccgact acacaaatca
     2041 gcgatttcca tgttgccact cgctttaatg atgatttctc ccgcgctgta ctggaggctg
     2101 aagtccaggt aagtttaaac aggatcttac taactaacat gctaacactg aattttcaga
     2161 tgtgcggcga gttgcgtgac tacctacggg taacagtttc tttatggcag ggtgaaacgc
     2221 aggtcgccag cggcaccgcg cctttcggcg gtgaaattat cgatgagcgt ggtggttatg
     2281 ccgatcgcgt cacactacgt ctgaacgtcg aaaacccgaa actgtggagc gccgaaatcc
     2341 cgaatctcta tcgtgcggtg gttgaactgc acaccgccga cggcacgctg attgaagcag
     2401 aagcctgcga tgtcggtttc cgcgaggtgc ggattgaaaa tggtctgctg ctgctgaacg
     2461 gcaagccgtt gctgattcga ggcgttaacc gtcacgagca tcatcctctg catggtcagg
     2521 tcatggatga gcagacgatg gtgcaggatg taagtttaaa ctattcgtta ctaactaact
     2581 ttaaacattt aaattttcag atcctgctga tgaagcagaa caactttaac gccgtgcgct
     2641 gttcgcatta tccgaaccat ccgctgtggt acacgctgtg cgaccgctac ggcctgtatg
     2701 tggtggatga agccaatatt gaaacccacg gcatggtgcc aatgaatcgt ctgaccgatg
     2761 atccgcgctg gctaccggcg atgagcgaac gcgtaacgcg aatggtgcag cgcgatcgta
     2821 atcacccgag tgtgatcatc tggtcgctgg ggaatggtaa gtttaaacag ttgaatacta
     2881 actaacggag atctttgaaa ttttcagaat caggccacgg cgctaatcac gacgcgctgt
     2941 atcgctggat caaatctgtc gatccttccc gcccggtgca gtatgaaggc ggcggagccg
     3001 acaccacggc caccgatatt atttgcccga tgtacgcgcg cgtggatgaa gaccagccct
     3061 tcccggctgt gccgaaatgg tccatcaaaa aatggctttc gctacctgga gagacgcgcc
     3121 cgctgattct ttgcgaggta agtttaaaca gaactctact aactaacaca ttagatccta
     3181 attttcagta cgctcacgcg atgggcaaca gtcttggcgg tttcgctaaa tactggcagg
     3241 cgtttcgtca gtatccccgt ttacagggcg gcttcgtctg ggactgggtg gatcagtcgc
     3301 tgattaaata tgatgaaaac ggcaacccgt ggtcggctta cggcggtgat tttggcgata
     3361 cgccgaacga tcgccagttc tgtatgaacg gtctggtctt tgccgaccgc acgccgcatc
     3421 cagcgtaagt ttaaacaata acctactaac taacgtagat aatttaaatt ttcaggctga
     3481 cggaagcaaa acaccagcag cagtttttcc agttccgttt atccgggcaa accatcgaag
     3541 tgaccagcga atacctgttc cgtcatagcg ataacgagct cctgcactgg atggtggcgc
     3601 tggatggtaa gccgctggca agcggtgaag tgcctctgga tgtcgctcca caaggtaaac
     3661 agttgattga actgcctgaa ctaccgcagc cggagagcgc cgggcaactc tggctcacag
     3721 tacgcgtagt gcaaccgaac gcgaccgcat ggtcagaagc cgggcacatc agcgcatggc
     3781 agcagtggag gtaagtttaa acaagatcct actaactaac tctacattga tgaattttca
     3841 gactggcgga aaacctcagt gtgacgctcc ccgccgcgtc ccacgccatc ccgcatctga
     3901 ccaccagcga aatggatttt tgcatcgagc tgggtaataa gcgttggcaa tttaaccgcc
     3961 agtcaggctt tctttcacag atgtggattg gcgataaaaa acaactgctg acgccgctgc
     4021 gcgatcagtt cacccgtgca ccgctggata acgacattgg cgtaagtgaa gcgacccgca
     4081 ttgaccctaa cgcctgggtc ggtaagttta aacaaagttg tactaactaa cgaagatctt
     4141 gataattttc agaacgctgg aaggcggcgg gccattacca ggccgaagca gcgttgttgc
     4201 agtgcacggc agatacactt gctgatgcgg tgctgattac gaccgctcac gcgtggcagc
     4261 atcaggggaa aaccttattt atcagccgga aaacctaccg gattgatggt agtggtcaaa
     4321 tggcgattac cgttgatgtt gaagtggcga gcgatacacc gcatccggcg cggattggcc
     4381 tgaactgcca gctggcgcag gtagcagagc gggtaaactg gctcggatta gggccgcaag
     4441 aaaactatcc cgaccgcctt actgccgcct gttttgaccg ctgggatctg ccattgtcag
     4501 acatgtagta agtttaaact tgatagtact aactaacatg tttcatttaa attttcagta
     4561 ccccgtacgt cttcccgagc gaaaacggtc tgcgctgcgg gacgcgcgaa ttgaattatg
     4621 gcccacacca gtggcgcggc gacttccagt tcaacatcag ccgctacagt caacagcaac
     4681 tgatggaaac cagccatcgc catctgctgc acgcggaaga aggcacatgg ctgaatatcg
     4741 acggtttcca tatggggatt ggtggcgacg actcctggag cccgtcagta tcggcggaat
     4801 tccaactgag cgccggtcgc taccattacc aacttgtctg gtgtcaaaaa taataggggc
     4861 cgctgtcatc agagtaagtt taaactgagt tctactaact aacgagtaat atttaaattt
     4921 tcagcatctc gcgcccgtgc ctctgacttc taagtccaat tactcttcaa catccctaca
     4981 tgctctttct ccctgtgctc ccacccccta tttttgttat tatcaaaaaa cttctcttaa
     5041 tttctttgtt ttttagcttc ttttaagtca cctctaacaa tgaaattgtg tagattcaaa
     5101 aatagaatta attcgtaata aaaagtcgaa aaaaattgtg ctccctcccc ccattaataa
     5161 taattctatc ccaaaatcta cacaatgttc tgtgtacact tcttatgttt tttacttctg
     5221 ataaattttt ttgaaacatc atagaaaaaa ccgcacacaa aataccttat catatgttac
     5281 gtttcagttt atgaccgcaa tttttatttc ttcgcacgtc tgggcctctc atgacgtcaa
     5341 atcatgctca tcgtgaaaaa gttttggagt atttttggaa tttttcaatc aagtgaaagt
     5401 ttatgaaatt aattttcctg cttttgcttt ttggggtttc ccctattgtt tgtcaagatt
     5461 tcgaggacgg cgtttttctt gctaaaatca caagtattga tgagcacgat gcaagaaaga
     5521 tcggaagaag gtttgggttt gaggctcagt ggaaggtgag tagaagttga taatttgaaa
     5581 gtggagtagt gtctatgggg tttttgcctt aaatgacaga atacattccc aatataccaa
     5641 acataactgt ttcctactag tcggccgtac gggccctttc gtctcgcgcg tttcggtgat
     5701 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5761 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5821 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     5881 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat
     5941 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     6001 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     6061 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     6121 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     6181 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     6241 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     6301 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     6361 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     6421 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     6481 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6541 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6601 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6661 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6721 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6781 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6841 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6901 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6961 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     7021 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     7081 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     7141 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     7201 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     7261 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     7321 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     7381 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     7441 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     7501 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     7561 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7621 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7681 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7741 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7801 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7861 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7921 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     7981 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     8041 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     8101 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     8161 gtaagtttaa acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa
     8221 atggctgaaa tcactcacaa cgatggatac gctaacaact tggaaatgaa at
//
