LOCUS       sequence-227-exp        6322 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 6322)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6322)
  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: L4026
FEATURES             Location/Qualifiers
     source          1..6322
                     /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"
     CDS             1797..1817
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1839..1859
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             1881..1901
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(1944..1972)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2720..2740
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3762..3784)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3884..3903
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3981..3999)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4067..4171
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4172..4240,4241..5032)
                     /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(4390..4409)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5203..5791
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5692..5711
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5945..5962
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6079..6100
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6115..6120,6121..6138,6139..6145)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6153..6169
                     /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     6158..6180
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6177..6193
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6177..6193
                     /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 aggtaccggc agaaaaaatg actgctccaa
     1801 agaagaagcg taaggtaccg gcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac
     1861 cggcagaaaa aatgactgct ccaaagaaga agcgtaaggt accggtagaa aaaatgagta
     1921 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
     1981 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
     2041 cccttaaatt tatttgcact actggaaaac tacctgttcc atgggtaagt ttaaacatat
     2101 atatactaac taaccctgat tatttaaatt ttcagccaac acttgtcact actttctgtt
     2161 atggtgttca atgcttctcg agatacccag atcatatgaa acggcatgac tttttcaaga
     2221 gtgccatgcc cgaaggttat gtacaggaaa gaactatatt tttcaaagat gacgggaact
     2281 acaagacacg taagtttaaa cagttcggta ctaactaacc atacatattt aaattttcag
     2341 gtgctgaagt caagtttgaa ggtgataccc ttgttaatag aatcgagtta aaaggtattg
     2401 attttaaaga agatggaaac attcttggac acaaattgga atacaactat aactcacaca
     2461 atgtatacat catggcagac aaacaaaaga atggaatcaa agttgtaagt ttaaacatga
     2521 ttttactaac taactaatct gatttaaatt ttcagaactt caaaattaga cacaacattg
     2581 aagatggaag cgttcaacta gcagaccatt atcaacaaaa tactccaatt ggcgatggcc
     2641 ctgtcctttt accagacaac cattacctgt ccacacaatc tgccctttcg aaagatccca
     2701 acgaaaagag agaccacatg gtccttcttg agtttgtaac agctgctggg attacacatg
     2761 gcatggatga actatacaaa tagcattcgt agaattgtaa gtttaaactt tctcatacta
     2821 actaacgcgc tctatttaaa ttttcagaat tccaactgag cgccggtcgc taccattacc
     2881 aacttgtctg gtgtcaaaaa taataggggc cgctgtcatc agagtaagtt taaactgagt
     2941 tctactaact aacgagtaat atttaaattt tcagcatctc gcgcccgtgc ctctgacttc
     3001 taagtccaat tactcttcaa catccctaca tgctctttct ccctgtgctc ccacccccta
     3061 tttttgttat tatcaaaaaa cttctcttaa tttctttgtt ttttagcttc ttttaagtca
     3121 cctctaacaa tgaaattgtg tagattcaaa aatagaatta attcgtaata aaaagtcgaa
     3181 aaaaattgtg ctccctcccc ccattaataa taattctatc ccaaaatcta cacaatgttc
     3241 tgtgtacact tcttatgttt tttacttctg ataaattttt ttgaaacatc atagaaaaaa
     3301 ccgcacacaa aataccttat catatgttac gtttcagttt atgaccgcaa tttttatttc
     3361 ttcgcacgtc tgggcctctc atgacgtcaa atcatgctca tcgtgaaaaa gttttggagt
     3421 atttttggaa tttttcaatc aagtgaaagt ttatgaaatt aattttcctg cttttgcttt
     3481 ttggggtttc ccctattgtt tgtcaagatt tcgaggacgg cgtttttctt gctaaaatca
     3541 caagtattga tgagcacgat gcaagaaaga tcggaagaag gtttgggttt gaggctcagt
     3601 ggaaggtgag tagaagttga taatttgaaa gtggagtagt gtctatgggg tttttgcctt
     3661 aaatgacaga atacattccc aatataccaa acataactgt ttcctactag tcggccgtac
     3721 gggccctttc gtctcgcgcg tttcggtgat gacggtgaaa acctctgaca catgcagctc
     3781 ccggagacgg tcacagcttg tctgtaagcg gatgccggga gcagacaagc ccgtcagggc
     3841 gcgtcagcgg gtgttggcgg gtgtcggggc tggcttaact atgcggcatc agagcagatt
     3901 gtactgagag tgcaccatat gcggtgtgaa ataccgcaca gatgcgtaag gagaaaatac
     3961 cgcatcaggc ggccttaagg gcctcgtgat acgcctattt ttataggtta atgtcatgat
     4021 aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgcgcg gaacccctat
     4081 ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat aaccctgata
     4141 aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc gtgtcgccct
     4201 tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa cgctggtgaa
     4261 agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac tggatctcaa
     4321 cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga tgagcacttt
     4381 taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag agcaactcgg
     4441 tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca cagaaaagca
     4501 tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca tgagtgataa
     4561 cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa ccgctttttt
     4621 gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc tgaatgaagc
     4681 cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa cgttgcgcaa
     4741 actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag actggatgga
     4801 ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct ggtttattgc
     4861 tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac tggggccaga
     4921 tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa ctatggatga
     4981 acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt aactgtcaga
     5041 ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat ttaaaaggat
     5101 ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg agttttcgtt
     5161 ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc ctttttttct
     5221 gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg tttgtttgcc
     5281 ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag cgcagatacc
     5341 aaatactgtc cttctagtgt agccgtagtt aggccaccac ttcaagaact ctgtagcacc
     5401 gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg gcgataagtc
     5461 gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc ggtcgggctg
     5521 aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg aactgagata
     5581 cctacagcgt gagcattgag aaagcgccac gcttcccgaa gggagaaagg cggacaggta
     5641 tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag ggggaaacgc
     5701 ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc gatttttgtg
     5761 atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct ttttacggtt
     5821 cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc ctgattctgt
     5881 ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc gaacgaccga
     5941 gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac cgcctctccc
     6001 cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact ggaaagcggg
     6061 cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc aggctttaca
     6121 ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat ttcacacagg
     6181 aaacagctat gaccatgatt acgccaagct gtaagtttaa acatgatctt actaactaac
     6241 tattctcatt taaattttca gagcttaaaa atggctgaaa tcactcacaa cgatggatac
     6301 gctaacaact tggaaatgaa at
//
