LOCUS       sequence-230-exp        7732 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 7732)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 7732)
  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: L4038
FEATURES             Location/Qualifiers
     source          1..7732
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3629..3649
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3671..3691
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3713..3733
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3755..3775
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3818..3846)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4594..4614
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5172..5194)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5294..5313
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5391..5409)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5477..5581
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5582..5650,5651..6442)
                     /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(5800..5819)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6613..7201
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7102..7121
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7355..7372
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7489..7510
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7525..7530,7531..7548,7549..7555)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7563..7579
                     /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     7568..7590
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7587..7603
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7587..7603
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttctgg agattgctcc agcagctctt aacagtgtct acgaccgtct tcgtgcaatt
       61 ctcatggaaa ctgaaagatc ggaaaatgca ctggatcgac gtattcagta tatgattgag
      121 actgcaatgc agattcgaaa ggacaaattt gcggtaaggt agaatatata aatagtttat
      181 tagaaaaaaa taaattagaa taatttaaat tcctactagc caatcaggcg acctttttgc
      241 gcatagttct attattgaaa aatttggaga atttctcata ttctcgctcg gaaatctgga
      301 attcgacgag atcttctggc ttctgtgcag ctgcatcgct ttgtgctccc tttctcgctt
      361 gtcttctgtg tacaccaaga accttgttga gttcatcaac tgaatctgtg actggcttgt
      421 tgctcactgg atgcactaga cgactgattc tcgagaaatc agattgagtt gcgattaggg
      481 tgacctagaa attgggaata atacgaactt ttgaaaatat tcaggaggat taaaaaaatt
      541 attctcgaca atcctacaaa tttacttatt gcaccatgtt gctccaacat ttttcattaa
      601 aagttaatga aaaaatgtag aaaatcggaa attggcaatt ttcagaccat ttttaagcat
      661 tttcaaaaaa aaattgcagc tgaaataaat gtcattttca gataaatcga gcgattttct
      721 gttgtctgac actagttttt agttttaaaa aatgttggaa gaacatggtg caataggtaa
      781 tttcatagaa tttccatgtg ttttttttca attaaccaat tatccaaatc ttccaaactc
      841 acattttgcg gagctgggct atcaagaatc tgctgcagtt ttataagacg agcatctctg
      901 atatcactga aaattaattt ttaatcaaaa cttgaatatc aactaaaccc acttattaac
      961 tttctcgatc ttctgtcgtt cggtacgatg acggtgaaga agccaattgt agtagttgat
     1021 ttggttcaag tcctttcggt gttgtacgtc agtgtcctgc aatgctattt agttataact
     1081 taggcctaag attcaattta atgaagtgat taaatttgtt ctctgaacct cttaagatga
     1141 tcttttggat tagaaacata taagacaggt ttacctatct attaaaaaac agatcaaaat
     1201 agatacgacc aaatcggata atccatgcct acctggcatc taggaacgtg ttcttagaag
     1261 atttcttacg taatcgtatg aagaaataac aatttgatcg ttggccagca aaaatagggt
     1321 tttaagtggg atagtgtttt tattagctaa ccggaaaatt ttatagtttt tttttgcaag
     1381 aaaccactga aaacccccta attgtataca ttttttggag cagcttctgg tctttttgag
     1441 caataaaatt cgataaaaca gaatttaagt gtaaattgtt cacatttagt ttctatttta
     1501 tcaaattttg ttgctcaaaa acattcgaag ctgctctaaa aaaatgcatt aaaaaagggg
     1561 ttttcagtgg tttttcacat taaaaaagct aattttaact aaaaatccat catatttcca
     1621 actttgtcac aacaataaaa tgctggtcaa aatgtgttcg aaaaaatgtt ttttttttta
     1681 atttttataa tttaaaaata gttttctttc gctgggacac atacattttt gggcgtaaat
     1741 tttcagttca aatttccatt tttacaacca taatcataaa gctacgtctg atctctctcg
     1801 cacttacctg cgcctgattc gaaagaacaa ccgtagccaa aagaacaaga agaacaagca
     1861 cgtagttgtg gtagtggacg ttcatcacgc aatactgacc aatggtcgtg gggtctcact
     1921 ttccgtacta ttgagagagg ggagactgaa gatggcaatt gaggacagtg tcttcgacgc
     1981 acgcatgcat ccataagcat aatccaggag ggatggagag aaaaatcttg tttctaagcc
     2041 cctccctttg taatacatac acatatctaa taccgaagaa tggctaattg aatggacgtc
     2101 agctgttgct gtagttgcca aggcatcatc gatgaaataa ctgaaagaaa gaattaaata
     2161 attattgcag gcgtatccgg cggtcattga agacttggac ttgattgagg aggaggatca
     2221 gatcatccat acacttaatt tggaggatgc ggttgatccg gaaaatgggc ttagtaagtg
     2281 actgaccaca cgcggggggc attaatttaa taaattgaat tccatttcag atgtgttcaa
     2341 actagatcca gaattcgaaa agaacgagga ggtttatgag gagatccgta aggaaatcat
     2401 tggaaacgcc gatatttcgg atgaggatgg tggcgacgag ttggatgatg aagaagaggg
     2461 tagtgatgtg gaagaggctc cgaagaagac tacagagatt attgataata ctgatcagaa
     2521 ttgactgctt tcagaaggta ttcattttga gttttgggcc ggcaaatctg taagttgccg
     2581 gttgccgaaa atttgctgaa tttgccggaa aaaaaaattc cggaatttat ttaaaaactt
     2641 tttgtaaaaa ttaaattaaa tttgcaactt ttcagagaag tctacctgac aatgcaatca
     2701 tctttggact accaagaagc tgctcacaaa ttgctgaaaa tgaagattcc agacagcatg
     2761 caggtcagcg atgttgcaaa gaaaaatttt cgaccaaaaa aaccaaccaa tcataaaatt
     2821 taaaaaaaaa ctccgttttt ttcttttttt ttatacgaga aaaaccaaaa aaatgtattt
     2881 ttgccaaatt ctaaaatact atccccgaaa ttttcaatat tttctctttc agaacgaact
     2941 ctgcgcgatg cttgtcgatt gttgtgctca acagcgtacc tacgagcgat tctacggaat
     3001 gctgagaaca cgttaaacac gaagatcata tttattttga gacccggatg ctctgaaaat
     3061 gtctgacata gatttaaaaa agcatatata tatttttcat tttcaacgtg aaagttttgt
     3121 gcaactttat agaatctcct attggcacat tgttttttat ttaactgagg cagtttttga
     3181 acaccttttt gaaactttga atctctttga agtatactgt cgaaaagact gacttgagcg
     3241 ttcgaaatgc cagaagaaaa ctatatttga atctcgcgct aaattgagaa atgcaaccgc
     3301 gctccactgg acaattggaa aaaaaattta ttcggaggcg acaacggtat tttcgaaatt
     3361 gattttctgt gtattttctc attttttata aattcttctt tgatttatcg ttcgtttgtg
     3421 agaaatttaa ttgtattcaa acttttttat agtaagatac cggtggtacc gctagccgta
     3481 cgaacccggg ttctagagga tcctatcgat tcgcggccgc tgtacacccc caaaggaccc
     3541 aaaggtatgt ttcgaatgat actaacataa catagaacat tttcaggagg acccttggag
     3601 ggtaccgagc tcagaaaaaa tgactgctcc aaagaagaag cgtaaggtac cggcagaaaa
     3661 aatgactgct ccaaagaaga agcgtaaggt accggcagaa aaaatgactg ctccaaagaa
     3721 gaagcgtaag gtaccggcag aaaaaatgac tgctccaaag aagaagcgta aggtaccggt
     3781 agaaaaaatg agtaaaggag aagaactttt cactggagtt gtcccaattc ttgttgaatt
     3841 agatggtgat gttaatgggc acaaattttc tgtcagtgga gagggtgaag gtgatgcaac
     3901 atacggaaaa cttaccctta aatttatttg cactactgga aaactacctg ttccatgggt
     3961 aagtttaaac atatatatac taactaaccc tgattattta aattttcagc caacacttgt
     4021 cactactttc tgttatggtg ttcaatgctt ctcgagatac ccagatcata tgaaacggca
     4081 tgactttttc aagagtgcca tgcccgaagg ttatgtacag gaaagaacta tatttttcaa
     4141 agatgacggg aactacaaga cacgtaagtt taaacagttc ggtactaact aaccatacat
     4201 atttaaattt tcaggtgctg aagtcaagtt tgaaggtgat acccttgtta atagaatcga
     4261 gttaaaaggt attgatttta aagaagatgg aaacattctt ggacacaaat tggaatacaa
     4321 ctataactca cacaatgtat acatcatggc agacaaacaa aagaatggaa tcaaagttgt
     4381 aagtttaaac atgattttac taactaacta atctgattta aattttcaga acttcaaaat
     4441 tagacacaac attgaagatg gaagcgttca actagcagac cattatcaac aaaatactcc
     4501 aattggcgat ggccctgtcc ttttaccaga caaccattac ctgtccacac aatctgccct
     4561 ttcgaaagat cccaacgaaa agagagacca catggtcctt cttgagtttg taacagctgc
     4621 tgggattaca catggcatgg acgaactata caaaagccca caagctttgg aattcgctag
     4681 ccggccatac aagtaatccg gatgatcgac gccaacgtcg ttgaattttc aaattttaaa
     4741 tactgaatat ttgttttttt tcctattatt tatttattct ctttgtgttt tttttcttgc
     4801 tttctaaaaa attaattcaa tccaaatcta aacatttttt tttctctttc cgtctcccaa
     4861 ttcgtattcc gctcctctca tctgaacaca atgtgcaagt ttatttatct tctcgctttc
     4921 atttcattag gacgtggggg gaattggtgg aagggggaaa cacacaaaag gatgatggaa
     4981 atgaaataag gacacacaat atgcaacaac attcaattca gaaatatgga ggaaggttta
     5041 aaagaaaaca taaaaatata tagaggagga aggaaaacta gtaaaaaata agcaaagaaa
     5101 ttaggcgaac gatgagaatt gtcctcgctt gggccctttc gtctcgcgcg tttcggtgat
     5161 gacggtgaaa acctctgaca catgcagctc ccggagacgg tcacagcttg tctgtaagcg
     5221 gatgccggga gcagacaagc ccgtcagggc gcgtcagcgg gtgttggcgg gtgtcggggc
     5281 tggcttaact atgcggcatc agagcagatt gtactgagag tgcaccatat gcggtgtgaa
     5341 ataccgcaca gatgcgtaag gagaaaatac cgcatcaggc ggccttaagg gcctcgtgat
     5401 acgcctattt ttataggtta atgtcatgat aataatggtt tcttagacgt caggtggcac
     5461 ttttcgggga aatgtgcgcg gaacccctat ttgtttattt ttctaaatac attcaaatat
     5521 gtatccgctc atgagacaat aaccctgata aatgcttcaa taatattgaa aaaggaagag
     5581 tatgagtatt caacatttcc gtgtcgccct tattcccttt tttgcggcat tttgccttcc
     5641 tgtttttgct cacccagaaa cgctggtgaa agtaaaagat gctgaagatc agttgggtgc
     5701 acgagtgggt tacatcgaac tggatctcaa cagcggtaag atccttgaga gttttcgccc
     5761 cgaagaacgt tttccaatga tgagcacttt taaagttctg ctatgtggcg cggtattatc
     5821 ccgtattgac gccgggcaag agcaactcgg tcgccgcata cactattctc agaatgactt
     5881 ggttgagtac tcaccagtca cagaaaagca tcttacggat ggcatgacag taagagaatt
     5941 atgcagtgct gccataacca tgagtgataa cactgcggcc aacttacttc tgacaacgat
     6001 cggaggaccg aaggagctaa ccgctttttt gcacaacatg ggggatcatg taactcgcct
     6061 tgatcgttgg gaaccggagc tgaatgaagc cataccaaac gacgagcgtg acaccacgat
     6121 gcctgtagca atggcaacaa cgttgcgcaa actattaact ggcgaactac ttactctagc
     6181 ttcccggcaa caattaatag actggatgga ggcggataaa gttgcaggac cacttctgcg
     6241 ctcggccctt ccggctggct ggtttattgc tgataaatct ggagccggtg agcgtgggtc
     6301 tcgcggtatc attgcagcac tggggccaga tggtaagccc tcccgtatcg tagttatcta
     6361 cacgacgggg agtcaggcaa ctatggatga acgaaataga cagatcgctg agataggtgc
     6421 ctcactgatt aagcattggt aactgtcaga ccaagtttac tcatatatac tttagattga
     6481 tttaaaactt catttttaat ttaaaaggat ctaggtgaag atcctttttg ataatctcat
     6541 gaccaaaatc ccttaacgtg agttttcgtt ccactgagcg tcagaccccg tagaaaagat
     6601 caaaggatct tcttgagatc ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa
     6661 accaccgcta ccagcggtgg tttgtttgcc ggatcaagag ctaccaactc tttttccgaa
     6721 ggtaactggc ttcagcagag cgcagatacc aaatactgtc cttctagtgt agccgtagtt
     6781 aggccaccac ttcaagaact ctgtagcacc gcctacatac ctcgctctgc taatcctgtt
     6841 accagtggct gctgccagtg gcgataagtc gtgtcttacc gggttggact caagacgata
     6901 gttaccggat aaggcgcagc ggtcgggctg aacggggggt tcgtgcacac agcccagctt
     6961 ggagcgaacg acctacaccg aactgagata cctacagcgt gagcattgag aaagcgccac
     7021 gcttcccgaa gggagaaagg cggacaggta tccggtaagc ggcagggtcg gaacaggaga
     7081 gcgcacgagg gagcttccag ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg
     7141 ccacctctga cttgagcgtc gatttttgtg atgctcgtca ggggggcgga gcctatggaa
     7201 aaacgccagc aacgcggcct ttttacggtt cctggccttt tgctggcctt ttgctcacat
     7261 gttctttcct gcgttatccc ctgattctgt ggataaccgt attaccgcct ttgagtgagc
     7321 tgataccgct cgccgcagcc gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga
     7381 agagcgccca atacgcaaac cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg
     7441 gcacgacagg tttcccgact ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta
     7501 gctcactcat taggcacccc aggctttaca ctttatgctt ccggctcgta tgttgtgtgg
     7561 aattgtgagc ggataacaat ttcacacagg aaacagctat gaccatgatt acgccaagct
     7621 gtaagtttaa acatgatctt actaactaac tattctcatt taaattttca gagcttaaaa
     7681 atggctgaaa tcactcacaa cgatggatac gctaacaact tggaaatgaa at
//
