LOCUS       sequence-143-exp        9720 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 9720)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 9720)
  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: L3137
FEATURES             Location/Qualifiers
     source          1..9720
                     /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(2747..2769)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(7160..7182)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     7282..7301
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(7379..7397)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        7465..7569
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(7570..7638,7639..8430)
                     /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(7788..7807)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      8601..9189
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     9090..9109
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     9343..9360
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    9477..9498
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(9513..9518,9519..9536,9537..9543)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9551..9567
                     /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     9556..9578
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     9575..9591
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     9575..9591
                     /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 caaatgcccg gtgggtgaag accagaaaca gcacctcgaa ctgagccgcg
     2701 atattgccca gcgtttcaac gcgctgtatg gcgagatcga tcccgtcgtt ttacaacgtc
     2761 gtgactggga aaaccctggc gttacccaac ttaatcgcct tgcagcacat ccccctttcg
     2821 ccagctggcg taatagcgaa gaggcccgca ccgatcgccc ttcccaacag ttgcgaaggt
     2881 aagtttaaac agatccatac taactaactt gttctgacat aattttcagc ttgaatggcg
     2941 aatggcgctt tgcctggttt ccggcaccag aagcggtgcc ggaaagctgg ctggagtgcg
     3001 atcttcctga ggccgatact gtcgtcgtcc cctcaaactg gcagatgcac ggttacgatg
     3061 cgcccatcta caccaacgtg acctatccca ttacggtcaa tccgccgttt gttcccacgg
     3121 agaatccgac gggttgttac tcgctcacat ttaatgttga tgaaagctgg ctacaggaag
     3181 gccagacgcg aattattttt gatggcgtta actcggcgtt tcatctgtgg tgcaacgggc
     3241 gctgggtcgg ttacggccag gtaagtttaa ttaagttgat actaactaac aagatctgat
     3301 taattttcag gacagtcgtt tgccgtctga atttgacctg agcgcatttt tacgcgccgg
     3361 agaaaaccgc ctcgcggtga tggtgctgcg ctggagtgac ggcagttatc tggaagatca
     3421 ggatatgtgg cggatgagcg gcattttccg tgacgtctcg ttgctgcata aaccgactac
     3481 acaaatcagc gatttccatg ttgccactcg ctttaatgat gatttctccc gcgctgtact
     3541 ggaggctgaa gtccaggtaa gtttaaacag gatcttacta actaacatgc taacactgaa
     3601 ttttcagatg tgcggcgagt tgcgtgacta cctacgggta acagtttctt tatggcaggg
     3661 tgaaacgcag gtcgccagcg gcaccgcgcc tttcggcggt gaaattatcg atgagcgtgg
     3721 tggttatgcc gatcgcgtca cactacgtct gaacgtcgaa aacccgaaac tgtggagcgc
     3781 cgaaatcccg aatctctatc gtgcggtggt tgaactgcac accgccgacg gcacgctgat
     3841 tgaagcagaa gcctgcgatg tcggtttccg cgaggtgcgg attgaaaatg gtctgctgct
     3901 gctgaacggc aagccgttgc tgattcgagg cgttaaccgt cacgagcatc atcctctgca
     3961 tggtcaggtc atggatgagc agacgatggt gcaggatgta agtttaaact attcgttact
     4021 aactaacttt aaacatttaa attttcagat cctgctgatg aagcagaaca actttaacgc
     4081 cgtgcgctgt tcgcattatc cgaaccatcc gctgtggtac acgctgtgcg accgctacgg
     4141 cctgtatgtg gtggatgaag ccaatattga aacccacggc atggtgccaa tgaatcgtct
     4201 gaccgatgat ccgcgctggc taccggcgat gagcgaacgc gtaacgcgaa tggtgcagcg
     4261 cgatcgtaat cacccgagtg tgatcatctg gtcgctgggg aatggtaagt ttaaacagtt
     4321 gaatactaac taacggagat ctttgaaatt ttcagaatca ggccacggcg ctaatcacga
     4381 cgcgctgtat cgctggatca aatctgtcga tccttcccgc ccggtgcagt atgaaggcgg
     4441 cggagccgac accacggcca ccgatattat ttgcccgatg tacgcgcgcg tggatgaaga
     4501 ccagcccttc ccggctgtgc cgaaatggtc catcaaaaaa tggctttcgc tacctggaga
     4561 gacgcgcccg ctgattcttt gcgaggtaag tttaaacaga actctactaa ctaacacatt
     4621 agatcctaat tttcagtacg ctcacgcgat gggcaacagt cttggcggtt tcgctaaata
     4681 ctggcaggcg tttcgtcagt atccccgttt acagggcggc ttcgtctggg actgggtgga
     4741 tcagtcgctg attaaatatg atgaaaacgg caacccgtgg tcggcttacg gcggtgattt
     4801 tggcgatacg ccgaacgatc gccagttctg tatgaacggt ctggtctttg ccgaccgcac
     4861 gccgcatcca gcgtaagttt aaacaataac ctactaacta acgtagataa tttaaatttt
     4921 caggctgacg gaagcaaaac accagcagca gtttttccag ttccgtttat ccgggcaaac
     4981 catcgaagtg accagcgaat acctgttccg tcatagcgat aacgagctcc tgcactggat
     5041 ggtggcgctg gatggtaagc cgctggcaag cggtgaagtg cctctggatg tcgctccaca
     5101 aggtaaacag ttgattgaac tgcctgaact accgcagccg gagagcgccg ggcaactctg
     5161 gctcacagta cgcgtagtgc aaccgaacgc gaccgcatgg tcagaagccg ggcacatcag
     5221 cgcatggcag cagtggaggt aagtttaaac aagatcctac taactaactc tacattgatg
     5281 aattttcaga ctggcggaaa acctcagtgt gacgctcccc gccgcgtccc acgccatccc
     5341 gcatctgacc accagcgaaa tggatttttg catcgagctg ggtaataagc gttggcaatt
     5401 taaccgccag tcaggctttc tttcacagat gtggattggc gataaaaaac aactgctgac
     5461 gccgctgcgc gatcagttca cccgtgcacc gctggataac gacattggcg taagtgaagc
     5521 gacccgcatt gaccctaacg cctgggtcgg taagtttaaa caaagttgta ctaactaacg
     5581 aagatcttga taattttcag aacgctggaa ggcggcgggc cattaccagg ccgaagcagc
     5641 gttgttgcag tgcacggcag atacacttgc tgatgcggtg ctgattacga ccgctcacgc
     5701 gtggcagcat caggggaaaa ccttatttat cagccggaaa acctaccgga ttgatggtag
     5761 tggtcaaatg gcgattaccg ttgatgttga agtggcgagc gatacaccgc atccggcgcg
     5821 gattggcctg aactgccagc tggcgcaggt agcagagcgg gtaaactggc tcggattagg
     5881 gccgcaagaa aactatcccg accgccttac tgccgcctgt tttgaccgct gggatctgcc
     5941 attgtcagac atgtagtaag tttaaacttg atagtactaa ctaacatgtt tcatttaaat
     6001 tttcagtacc ccgtacgtct tcccgagcga aaacggtctg cgctgcggga cgcgcgaatt
     6061 gaattatggc ccacaccagt ggcgcggcga cttccagttc aacatcagcc gctacagtca
     6121 acagcaactg atggaaacca gccatcgcca tctgctgcac gcggaagaag gcacatggct
     6181 gaatatcgac ggtttccata tggggattgg tggcgacgac tcctggagcc cgtcagtatc
     6241 ggcggaattc caactgagcg ccggtcgcta ccattaccaa cttgtctggt gtcaaaaata
     6301 ataggggccg ctgtcatcag agtaagttta aactgagttc tactaactaa cgagtaatat
     6361 ttaaattttc agcatctcgc gcccgtgcct ctgacttcta agtccaatta ctcttcaaca
     6421 tccctacatg ctctttctcc ctgtgctccc accccctatt tttgttatta tcaaaaaact
     6481 tctcttaatt tctttgtttt ttagcttctt ttaagtcacc tctaacaatg aaattgtgta
     6541 gattcaaaaa tagaattaat tcgtaataaa aagtcgaaaa aaattgtgct ccctcccccc
     6601 attaataata attctatccc aaaatctaca caatgttctg tgtacacttc ttatgttttt
     6661 tacttctgat aaattttttt gaaacatcat agaaaaaacc gcacacaaaa taccttatca
     6721 tatgttacgt ttcagtttat gaccgcaatt tttatttctt cgcacgtctg ggcctctcat
     6781 gacgtcaaat catgctcatc gtgaaaaagt tttggagtat ttttggaatt tttcaatcaa
     6841 gtgaaagttt atgaaattaa ttttcctgct tttgcttttt ggggtttccc ctattgtttg
     6901 tcaagatttc gaggacggcg tttttcttgc taaaatcaca agtattgatg agcacgatgc
     6961 aagaaagatc ggaagaaggt ttgggtttga ggctcagtgg aaggtgagta gaagttgata
     7021 atttgaaagt ggagtagtgt ctatggggtt tttgccttaa atgacagaat acattcccaa
     7081 tataccaaac ataactgttt cctactagtc ggccgtacgg gccctttcgt ctcgcgcgtt
     7141 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     7201 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     7261 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     7321 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     7381 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     7441 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     7501 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     7561 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     7621 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     7681 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     7741 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     7801 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     7861 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     7921 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     7981 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     8041 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     8101 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     8161 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     8221 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     8281 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     8341 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     8401 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     8461 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     8521 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     8581 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     8641 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     8701 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     8761 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     8821 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     8881 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     8941 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     9001 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     9061 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     9121 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     9181 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     9241 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     9301 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     9361 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     9421 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     9481 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     9541 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     9601 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     9661 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
