LOCUS       sequence-291-exp        7560 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 7560)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 7560)
  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: L4809
FEATURES             Location/Qualifiers
     source          1..7560
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             3493..3513
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3538..3558
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     CDS             3583..3603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(3646..3674)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     4422..4442
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(5000..5022)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5122..5141
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5219..5237)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5305..5409
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5410..5478,5479..6270)
                     /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(5628..5647)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6441..7029
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6930..6949
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7183..7200
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7317..7338
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7353..7358,7359..7376,7377..7383)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7391..7407
                     /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     7396..7418
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     7415..7431
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     7415..7431
                     /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 gagctcagaa
     3481 aaaatgaccg ccccaaagaa gaaacgcaaa gtaccgagct cagaaaaaat gaccgctcca
     3541 aagaagaaac gcaaagtacc gagctcagaa aaaatgacgg cacctaaaaa gaaacgcaaa
     3601 gtaccggtag aaaaaatgag taaaggagaa gaacttttca ctggagttgt cccaattctt
     3661 gttgaattag atggtgatgt taatgggcac aaattttctg tcagtggaga gggtgaaggt
     3721 gatgcaacat acggaaaact tacccttaaa tttatttgca ctactggaaa actacctgtt
     3781 ccatgggtaa gtttaaacat atatatacta actaaccctg attatttaaa ttttcagcca
     3841 acacttgtca ctactttctg ttatggtgtt caatgcttct cgagataccc agatcatatg
     3901 aaacggcatg actttttcaa gagtgccatg cccgaaggtt atgtacagga aagaactata
     3961 tttttcaaag atgacgggaa ctacaagaca cgtaagttta aacagttcgg tactaactaa
     4021 ccatacatat ttaaattttc aggtgctgaa gtcaagtttg aaggtgatac ccttgttaat
     4081 agaatcgagt taaaaggtat tgattttaaa gaagatggaa acattcttgg acacaaattg
     4141 gaatacaact ataactcaca caatgtatac atcatggcag acaaacaaaa gaatggaatc
     4201 aaagttgtaa gtttaaacat gattttacta actaactaat ctgatttaaa ttttcagaac
     4261 ttcaaaatta gacacaacat tgaagatgga agcgttcaac tagcagacca ttatcaacaa
     4321 aatactccaa ttggcgatgg ccctgtcctt ttaccagaca accattacct gtccacacaa
     4381 tctgcccttt cgaaagatcc caacgaaaag agagaccaca tggtccttct tgagtttgta
     4441 acagctgctg ggattacaca tggcatggac gaactataca aaagcccaca agctttggaa
     4501 ttcgctagcc ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa
     4561 attttaaata ctgaatattt gttttttttc ctattattta tttattctct ttgtgttttt
     4621 tttcttgctt tctaaaaaat taattcaatc caaatctaaa catttttttt tctctttccg
     4681 tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt atttatcttc
     4741 tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca cacaaaagga
     4801 tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga aatatggagg
     4861 aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt aaaaaataag
     4921 caaagaaatt aggcgaacga tgagaattgt cctcgcttgg gccctttcgt ctcgcgcgtt
     4981 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     5041 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     5101 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     5161 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     5221 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     5281 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     5341 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     5401 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     5461 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     5521 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     5581 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     5641 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     5701 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     5761 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     5821 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     5881 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     5941 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     6001 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     6061 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     6121 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     6181 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     6241 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     6301 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     6361 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     6421 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     6481 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     6541 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     6601 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     6661 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     6721 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     6781 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     6841 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     6901 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     6961 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     7021 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     7081 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     7141 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     7201 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     7261 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     7321 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     7381 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     7441 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     7501 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
