LOCUS       sequence-123-exp        6141 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 6141)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 6141)
  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: L2605
FEATURES             Location/Qualifiers
     source          1..6141
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1755..1775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1818..1846)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2594..2614
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3581..3603)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3703..3722
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3800..3818)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3886..3990
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3991..4059,4060..4851)
                     /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(4209..4228)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5022..5610
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5511..5530
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5764..5781
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5898..5919
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5934..5939,5940..5957,5958..5964)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5972..5988
                     /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     5977..5999
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5996..6012
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5996..6012
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggc cttggtcgag tatgagaaac aacattaaca atgttttggg
       61 agaaacatgt agttttgtac gtccggtcca gtgagagttt tcaacttatt ttggaagaaa
      121 tttttttttg acaaaatttt gcaatttccc acgcagaaat acggtatgcc ccgtatctcg
      181 ccacgaacat aatattttaa attgacggaa aggtgtgcgc ctttaagaag cacagtaacc
      241 attttccggt tttttttttg aaattgcctt tgatttttac ttttttctcc caaaaacgag
      301 aaatttaata taatccatca aaatctttgt gaacgagaga aatatagtga aaaactcaaa
      361 gaaaattgcg aaatggatta ctgtactcca taaaggcgca catcttttcg aaaaatcgtg
      421 aaaaattgca tctgaccaac agttttgcgt ggaatcttcc tagattcaaa tttcctatga
      481 gtcataaaac tatcatttta tcggacaata actacacgtg taccttgttt ccatatgttc
      541 taccaatata atttgtgttc atatgtggcc aggtacggga aattagaaca cggactgatt
      601 taggaaggtt caaattcgag aacaatggac gacttgctgc acgaaaacgt ggattattct
      661 gaaaccgcct atgtgatcac tatcgattta gcacttcggt tttctttttc cgttttcctt
      721 tatattccgt attgtatgcc gacaataacg ctttttcttt gtataaacat ttccagaagc
      781 agagcaaatt tgtttattcc agaattgact gaatattgaa ataatgttta cagacatact
      841 gtagattttt ttctattcaa acatgaatac agtttttcgt atttttaatc ttcagataag
      901 ttttgtgtgt gtttttgaga ataatattta aaaaagataa gcaaatatca gaactacggt
      961 aagcgaaggg gtacggtcgt caattgcgga cttccgagct gtgcacggcg ccaaaattta
     1021 ttttgagttt taaaatgatt ttctttgttt ttttttcttt ttttgacacg ataagaacat
     1081 ttttctgtat ttctaatatt ttaaatgaga tcgtaataag tttatttagc tccatcacat
     1141 tattttcgga atattcatcg cttgaaaacg gcgtaaaata ggttccaagt aaatcattgc
     1201 aaacaaatct ccgtttatcg ttctaacaaa gtttttcgtt cactttctta tgtaaatcgt
     1261 tttccccact tcttcttcac gggaatgatc aattgaaagg ccataaattg gttgttttct
     1321 ccacatttcg caaccaacat gcagctgtcg gataggcaat aaaataatgt aatcaatgag
     1381 ttttccgatt tatcctaaca gttctttttc tagtgttcct attcgaaaaa agtactgtta
     1441 tccttctttt tggaccgccc atctcaggtg tcaagacaat cgcaacacac tcagaaaatg
     1501 ggcggagtat gccacccggc cttcattcgt cgctccttgt cttctccatt ttcgtttttt
     1561 tcctgttctc cgttcttttt ttgtactttt gtgataatgc tgaagtgttc ttttttttcg
     1621 cttaaccgca tctcttgatt taattcatat tttctttcag attcatatac aataacacca
     1681 agagctgctc taacaactaa taataacaga atctagagga tccccgggta ccgagctcag
     1741 aaaaaatgac tgctccaaag aagaagcgta aggtaccggt agaaaaaatg agtaaaggag
     1801 aagaactttt cactggagtt gtcccaattc ttgttgaatt agatggtgat gttaatgggc
     1861 acaaattttc tgtcagtgga gagggtgaag gtgatgcaac atacggaaaa cttaccctta
     1921 aatttatttg cactactgga aaactacctg ttccatgggt aagtttaaac atatatatac
     1981 taactaaccc tgattattta aattttcagc caacacttgt cactactttc tgttatggtg
     2041 ttcaatgctt ctcgagatac ccagatcata tgaaacggca tgactttttc aagagtgcca
     2101 tgcccgaagg ttatgtacag gaaagaacta tatttttcaa agatgacggg aactacaaga
     2161 cacgtaagtt taaacagttc ggtactaact aaccatacat atttaaattt tcaggtgctg
     2221 aagtcaagtt tgaaggtgat acccttgtta atagaatcga gttaaaaggt attgatttta
     2281 aagaagatgg aaacattctt ggacacaaat tggaatacaa ctataactca cacaatgtat
     2341 acatcatggc agacaaacaa aagaatggaa tcaaagttgt aagtttaaac atgattttac
     2401 taactaacta atctgattta aattttcaga acttcaaaat tagacacaac attgaagatg
     2461 gaagcgttca actagcagac cattatcaac aaaatactcc aattggcgat ggccctgtcc
     2521 ttttaccaga caaccattac ctgtccacac aatctgccct ttcgaaagat cccaacgaaa
     2581 agagagacca catggtcctt cttgagtttg taacagctgc tgggattaca catggcatgg
     2641 atgaactata caaatagcat tcgtagaatt ccaactgagc gccggtcgct accattacca
     2701 acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt aaactgagtt
     2761 ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc tctgacttct
     2821 aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc caccccctat
     2881 ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct tttaagtcac
     2941 ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa aaagtcgaaa
     3001 aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac acaatgttct
     3061 gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca tagaaaaaac
     3121 cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat ttttatttct
     3181 tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag ttttggagta
     3241 tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc ttttgctttt
     3301 tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg ctaaaatcac
     3361 aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg aggctcagtg
     3421 gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt ttttgcctta
     3481 aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt cggccgtacg
     3541 ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac atgcagctcc
     3601 cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg
     3661 cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca gagcagattg
     3721 tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc
     3781 gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa tgtcatgata
     3841 ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg aacccctatt
     3901 tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata accctgataa
     3961 atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg tgtcgccctt
     4021 attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac gctggtgaaa
     4081 gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact ggatctcaac
     4141 agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat gagcactttt
     4201 aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga gcaactcggt
     4261 cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac agaaaagcat
     4321 cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat gagtgataac
     4381 actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac cgcttttttg
     4441 cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct gaatgaagcc
     4501 ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa
     4561 ctattaactg gcgaactact tactctagct tcccggcaac aattaataga ctggatggag
     4621 gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg gtttattgct
     4681 gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact ggggccagat
     4741 ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac tatggatgaa
     4801 cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta actgtcagac
     4861 caagtttact catatatact ttagattgat ttaaaacttc atttttaatt taaaaggatc
     4921 taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga gttttcgttc
     4981 cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc tttttttctg
     5041 cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg
     5101 gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc gcagatacca
     5161 aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc tgtagcaccg
     5221 cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg cgataagtcg
     5281 tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg gtcgggctga
     5341 acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga actgagatac
     5401 ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc ggacaggtat
     5461 ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg gggaaacgcc
     5521 tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg atttttgtga
     5581 tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt tttacggttc
     5641 ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc tgattctgtg
     5701 gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg aacgaccgag
     5761 cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc
     5821 gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg gaaagcgggc
     5881 agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca ggctttacac
     5941 tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt tcacacagga
     6001 aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta ctaactaact
     6061 attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac gatggatacg
     6121 ctaacaactt ggaaatgaaa t
//
