LOCUS       sequence-128-exp        7532 bp    DNA     circular SYN 15-APR-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7532)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 7532)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
COMMENT     Sequence Label: L2682
FEATURES             Location/Qualifiers
     source          1..7532
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             583..603
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(693..715)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(5106..5128)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5228..5247
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5325..5343)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5411..5515
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5516..5584,5585..6376)
                     /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(5734..5753)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6547..7135
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     7036..7055
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7289..7306
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7423..7444
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7459..7464,7465..7482,7483..7489)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7497..7513
                     /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     7502..7524
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7521..7532,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7521..7532,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tcaccaaaaa
       61 cggaacgttg agctggacgg aaatagtggt aaagtgacat gattatagtt tgaagatttc
      121 taatttcaca attagagcaa atgttgttcg gtatttattt tcaacggtat ttatactatt
      181 ttccaccttt ttctagaaca ttcgagctgc ttgttgcaaa aggagggcga ctcacattcg
      241 gtacatggaa aagtagtgta cacaataaag agacccagat acattttccg tctgcgtctc
      301 tttgcaccca ccgggagtat tttcaaacga atgcatctag gaccttctag aacattctgt
      361 aaggctgcag aatgcgggta tataaggaaa gcgggctcag aggaagccaa cacgctttgt
      421 tctagtgcat ctaaaaaact tcgaaaatcc tcatcggatc cccgggattg gccaaaggac
      481 ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga ggacccttgg
      541 ctagaggatc cccgggtacc gagctcagaa aaaatgactg ctccaaagaa gaagcgtaag
      601 gtaccggtgg gtgaagacca gaaacagcac ctcgaactga gccgcgatat tgcccagcgt
      661 ttcaacgcgc tgtatggcga gatcgatccc gtcgttttac aacgtcgtga ctgggaaaac
      721 cctggcgtta cccaacttaa tcgccttgca gcacatcccc ctttcgccag ctggcgtaat
      781 agcgaagagg cccgcaccga tcgcccttcc caacagttgc gaaggtaagt ttaaacagat
      841 ccatactaac taacttgttc tgacataatt ttcagcttga atggcgaatg gcgctttgcc
      901 tggtttccgg caccagaagc ggtgccggaa agctggctgg agtgcgatct tcctgaggcc
      961 gatactgtcg tcgtcccctc aaactggcag atgcacggtt acgatgcgcc catctacacc
     1021 aacgtgacct atcccattac ggtcaatccg ccgtttgttc ccacggagaa tccgacgggt
     1081 tgttactcgc tcacatttaa tgttgatgaa agctggctac aggaaggcca gacgcgaatt
     1141 atttttgatg gcgttaactc ggcgtttcat ctgtggtgca acgggcgctg ggtcggttac
     1201 ggccaggtaa gtttaattaa gttgatacta actaacaaga tctgattaat tttcaggaca
     1261 gtcgtttgcc gtctgaattt gacctgagcg catttttacg cgccggagaa aaccgcctcg
     1321 cggtgatggt gctgcgctgg agtgacggca gttatctgga agatcaggat atgtggcgga
     1381 tgagcggcat tttccgtgac gtctcgttgc tgcataaacc gactacacaa atcagcgatt
     1441 tccatgttgc cactcgcttt aatgatgatt tctcccgcgc tgtactggag gctgaagtcc
     1501 aggtaagttt aaacaggatc ttactaacta acatgctaac actgaatttt cagatgtgcg
     1561 gcgagttgcg tgactaccta cgggtaacag tttctttatg gcagggtgaa acgcaggtcg
     1621 ccagcggcac cgcgcctttc ggcggtgaaa ttatcgatga gcgtggtggt tatgccgatc
     1681 gcgtcacact acgtctgaac gtcgaaaacc cgaaactgtg gagcgccgaa atcccgaatc
     1741 tctatcgtgc ggtggttgaa ctgcacaccg ccgacggcac gctgattgaa gcagaagcct
     1801 gcgatgtcgg tttccgcgag gtgcggattg aaaatggtct gctgctgctg aacggcaagc
     1861 cgttgctgat tcgaggcgtt aaccgtcacg agcatcatcc tctgcatggt caggtcatgg
     1921 atgagcagac gatggtgcag gatgtaagtt taaactattc gttactaact aactttaaac
     1981 atttaaattt tcagatcctg ctgatgaagc agaacaactt taacgccgtg cgctgttcgc
     2041 attatccgaa ccatccgctg tggtacacgc tgtgcgaccg ctacggcctg tatgtggtgg
     2101 atgaagccaa tattgaaacc cacggcatgg tgccaatgaa tcgtctgacc gatgatccgc
     2161 gctggctacc ggcgatgagc gaacgcgtaa cgcgaatggt gcagcgcgat cgtaatcacc
     2221 cgagtgtgat catctggtcg ctggggaatg gtaagtttaa acagttgaat actaactaac
     2281 ggagatcttt gaaattttca gaatcaggcc acggcgctaa tcacgacgcg ctgtatcgct
     2341 ggatcaaatc tgtcgatcct tcccgcccgg tgcagtatga aggcggcgga gccgacacca
     2401 cggccaccga tattatttgc ccgatgtacg cgcgcgtgga tgaagaccag cccttcccgg
     2461 ctgtgccgaa atggtccatc aaaaaatggc tttcgctacc tggagagacg cgcccgctga
     2521 ttctttgcga ggtaagttta aacagaactc tactaactaa cacattagat cctaattttc
     2581 agtacgctca cgcgatgggc aacagtcttg gcggtttcgc taaatactgg caggcgtttc
     2641 gtcagtatcc ccgtttacag ggcggcttcg tctgggactg ggtggatcag tcgctgatta
     2701 aatatgatga aaacggcaac ccgtggtcgg cttacggcgg tgattttggc gatacgccga
     2761 acgatcgcca gttctgtatg aacggtctgg tctttgccga ccgcacgccg catccagcgt
     2821 aagtttaaac aataacctac taactaacgt agataattta aattttcagg ctgacggaag
     2881 caaaacacca gcagcagttt ttccagttcc gtttatccgg gcaaaccatc gaagtgacca
     2941 gcgaatacct gttccgtcat agcgataacg agctcctgca ctggatggtg gcgctggatg
     3001 gtaagccgct ggcaagcggt gaagtgcctc tggatgtcgc tccacaaggt aaacagttga
     3061 ttgaactgcc tgaactaccg cagccggaga gcgccgggca actctggctc acagtacgcg
     3121 tagtgcaacc gaacgcgacc gcatggtcag aagccgggca catcagcgca tggcagcagt
     3181 ggaggtaagt ttaaacaaga tcctactaac taactctaca ttgatgaatt ttcagactgg
     3241 cggaaaacct cagtgtgacg ctccccgccg cgtcccacgc catcccgcat ctgaccacca
     3301 gcgaaatgga tttttgcatc gagctgggta ataagcgttg gcaatttaac cgccagtcag
     3361 gctttctttc acagatgtgg attggcgata aaaaacaact gctgacgccg ctgcgcgatc
     3421 agttcacccg tgcaccgctg gataacgaca ttggcgtaag tgaagcgacc cgcattgacc
     3481 ctaacgcctg ggtcggtaag tttaaacaaa gttgtactaa ctaacgaaga tcttgataat
     3541 tttcagaacg ctggaaggcg gcgggccatt accaggccga agcagcgttg ttgcagtgca
     3601 cggcagatac acttgctgat gcggtgctga ttacgaccgc tcacgcgtgg cagcatcagg
     3661 ggaaaacctt atttatcagc cggaaaacct accggattga tggtagtggt caaatggcga
     3721 ttaccgttga tgttgaagtg gcgagcgata caccgcatcc ggcgcggatt ggcctgaact
     3781 gccagctggc gcaggtagca gagcgggtaa actggctcgg attagggccg caagaaaact
     3841 atcccgaccg ccttactgcc gcctgttttg accgctggga tctgccattg tcagacatgt
     3901 agtaagttta aacttgatag tactaactaa catgtttcat ttaaattttc agtaccccgt
     3961 acgtcttccc gagcgaaaac ggtctgcgct gcgggacgcg cgaattgaat tatggcccac
     4021 accagtggcg cggcgacttc cagttcaaca tcagccgcta cagtcaacag caactgatgg
     4081 aaaccagcca tcgccatctg ctgcacgcgg aagaaggcac atggctgaat atcgacggtt
     4141 tccatatggg gattggtggc gacgactcct ggagcccgtc agtatcggcg gaattccaac
     4201 tgagcgccgg tcgctaccat taccaacttg tctggtgtca aaaataatag gggccgctgt
     4261 catcagagta agtttaaact gagttctact aactaacgag taatatttaa attttcagca
     4321 tctcgcgccc gtgcctctga cttctaagtc caattactct tcaacatccc tacatgctct
     4381 ttctccctgt gctcccaccc cctatttttg ttattatcaa aaaacttctc ttaatttctt
     4441 tgttttttag cttcttttaa gtcacctcta acaatgaaat tgtgtagatt caaaaataga
     4501 attaattcgt aataaaaagt cgaaaaaaat tgtgctccct ccccccatta ataataattc
     4561 tatcccaaaa tctacacaat gttctgtgta cacttcttat gttttttact tctgataaat
     4621 ttttttgaaa catcatagaa aaaaccgcac acaaaatacc ttatcatatg ttacgtttca
     4681 gtttatgacc gcaattttta tttcttcgca cgtctgggcc tctcatgacg tcaaatcatg
     4741 ctcatcgtga aaaagttttg gagtattttt ggaatttttc aatcaagtga aagtttatga
     4801 aattaatttt cctgcttttg ctttttgggg tttcccctat tgtttgtcaa gatttcgagg
     4861 acggcgtttt tcttgctaaa atcacaagta ttgatgagca cgatgcaaga aagatcggaa
     4921 gaaggtttgg gtttgaggct cagtggaagg tgagtagaag ttgataattt gaaagtggag
     4981 tagtgtctat ggggtttttg ccttaaatga cagaatacat tcccaatata ccaaacataa
     5041 ctgtttccta ctagtcggcc gtacgggccc tttcgtctcg cgcgtttcgg tgatgacggt
     5101 gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta agcggatgcc
     5161 gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg gggctggctt
     5221 aactatgcgg catcagagca gattgtactg agagtgcacc atatgcggtg tgaaataccg
     5281 cacagatgcg taaggagaaa ataccgcatc aggcggcctt aagggcctcg tgatacgcct
     5341 atttttatag gttaatgtca tgataataat ggtttcttag acgtcaggtg gcacttttcg
     5401 gggaaatgtg cgcggaaccc ctatttgttt atttttctaa atacattcaa atatgtatcc
     5461 gctcatgaga caataaccct gataaatgct tcaataatat tgaaaaagga agagtatgag
     5521 tattcaacat ttccgtgtcg cccttattcc cttttttgcg gcattttgcc ttcctgtttt
     5581 tgctcaccca gaaacgctgg tgaaagtaaa agatgctgaa gatcagttgg gtgcacgagt
     5641 gggttacatc gaactggatc tcaacagcgg taagatcctt gagagttttc gccccgaaga
     5701 acgttttcca atgatgagca cttttaaagt tctgctatgt ggcgcggtat tatcccgtat
     5761 tgacgccggg caagagcaac tcggtcgccg catacactat tctcagaatg acttggttga
     5821 gtactcacca gtcacagaaa agcatcttac ggatggcatg acagtaagag aattatgcag
     5881 tgctgccata accatgagtg ataacactgc ggccaactta cttctgacaa cgatcggagg
     5941 accgaaggag ctaaccgctt ttttgcacaa catgggggat catgtaactc gccttgatcg
     6001 ttgggaaccg gagctgaatg aagccatacc aaacgacgag cgtgacacca cgatgcctgt
     6061 agcaatggca acaacgttgc gcaaactatt aactggcgaa ctacttactc tagcttcccg
     6121 gcaacaatta atagactgga tggaggcgga taaagttgca ggaccacttc tgcgctcggc
     6181 ccttccggct ggctggttta ttgctgataa atctggagcc ggtgagcgtg ggtctcgcgg
     6241 tatcattgca gcactggggc cagatggtaa gccctcccgt atcgtagtta tctacacgac
     6301 ggggagtcag gcaactatgg atgaacgaaa tagacagatc gctgagatag gtgcctcact
     6361 gattaagcat tggtaactgt cagaccaagt ttactcatat atactttaga ttgatttaaa
     6421 acttcatttt taatttaaaa ggatctaggt gaagatcctt tttgataatc tcatgaccaa
     6481 aatcccttaa cgtgagtttt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     6541 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     6601 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     6661 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     6721 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     6781 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     6841 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     6901 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     6961 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     7021 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     7081 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     7141 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     7201 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     7261 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     7321 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     7381 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatgtga gttagctcac
     7441 tcattaggca ccccaggctt tacactttat gcttccggct cgtatgttgt gtggaattgt
     7501 gagcggataa caatttcaca caggaaacag ct
//
