LOCUS       sequence-77-expo        7110 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 7110)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7110)
  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: pPD89_20
FEATURES             Location/Qualifiers
     source          1..7110
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             161..181
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(271..293)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4684..4706)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4806..4825
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4903..4921)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4989..5093
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5094..5162,5163..5954)
                     /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(5312..5331)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6125..6713
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6614..6633
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6867..6884
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7001..7022
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7037..7042,7043..7060,7061..7067)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7075..7091
                     /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     7080..7102
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7099..7110,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7099..7110,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctagaggatc cccgggattg
       61 gccaaaggac ccaaaggtat gtttcgaatg atactaacat aacatagaac attttcagga
      121 ggacccttgg agggtaccga gctcagaaaa aatgactgct ccaaagaaga agcgtaaggt
      181 accggtgggt gaagaccaga aacagcacct cgaactgagc cgcgatattg cccagcgttt
      241 caacgcgctg tatggcgaga tcgatcccgt cgttttacaa cgtcgtgact gggaaaaccc
      301 tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct ggcgtaatag
      361 cgaagaggcc cgcaccgatc gcccttccca acagttgcga aggtaagttt aaacagatcc
      421 atactaacta acttgttctg acataatttt cagcttgaat ggcgaatggc gctttgcctg
      481 gtttccggca ccagaagcgg tgccggaaag ctggctggag tgcgatcttc ctgaggccga
      541 tactgtcgtc gtcccctcaa actggcagat gcacggttac gatgcgccca tctacaccaa
      601 cgtgacctat cccattacgg tcaatccgcc gtttgttccc acggagaatc cgacgggttg
      661 ttactcgctc acatttaatg ttgatgaaag ctggctacag gaaggccaga cgcgaattat
      721 ttttgatggc gttaactcgg cgtttcatct gtggtgcaac gggcgctggg tcggttacgg
      781 ccaggtaagt ttaattaagt tgatactaac taacaagatc tgattaattt tcaggacagt
      841 cgtttgccgt ctgaatttga cctgagcgca tttttacgcg ccggagaaaa ccgcctcgcg
      901 gtgatggtgc tgcgctggag tgacggcagt tatctggaag atcaggatat gtggcggatg
      961 agcggcattt tccgtgacgt ctcgttgctg cataaaccga ctacacaaat cagcgatttc
     1021 catgttgcca ctcgctttaa tgatgatttc tcccgcgctg tactggaggc tgaagtccag
     1081 gtaagtttaa acaggatctt actaactaac atgctaacac tgaattttca gatgtgcggc
     1141 gagttgcgtg actacctacg ggtaacagtt tctttatggc agggtgaaac gcaggtcgcc
     1201 agcggcaccg cgcctttcgg cggtgaaatt atcgatgagc gtggtggtta tgccgatcgc
     1261 gtcacactac gtctgaacgt cgaaaacccg aaactgtgga gcgccgaaat cccgaatctc
     1321 tatcgtgcgg tggttgaact gcacaccgcc gacggcacgc tgattgaagc agaagcctgc
     1381 gatgtcggtt tccgcgaggt gcggattgaa aatggtctgc tgctgctgaa cggcaagccg
     1441 ttgctgattc gaggcgttaa ccgtcacgag catcatcctc tgcatggtca ggtcatggat
     1501 gagcagacga tggtgcagga tgtaagttta aactattcgt tactaactaa ctttaaacat
     1561 ttaaattttc agatcctgct gatgaagcag aacaacttta acgccgtgcg ctgttcgcat
     1621 tatccgaacc atccgctgtg gtacacgctg tgcgaccgct acggcctgta tgtggtggat
     1681 gaagccaata ttgaaaccca cggcatggtg ccaatgaatc gtctgaccga tgatccgcgc
     1741 tggctaccgg cgatgagcga acgcgtaacg cgaatggtgc agcgcgatcg taatcacccg
     1801 agtgtgatca tctggtcgct ggggaatggt aagtttaaac agttgaatac taactaacgg
     1861 agatctttga aattttcaga atcaggccac ggcgctaatc acgacgcgct gtatcgctgg
     1921 atcaaatctg tcgatccttc ccgcccggtg cagtatgaag gcggcggagc cgacaccacg
     1981 gccaccgata ttatttgccc gatgtacgcg cgcgtggatg aagaccagcc cttcccggct
     2041 gtgccgaaat ggtccatcaa aaaatggctt tcgctacctg gagagacgcg cccgctgatt
     2101 ctttgcgagg taagtttaaa cagaactcta ctaactaaca cattagatcc taattttcag
     2161 tacgctcacg cgatgggcaa cagtcttggc ggtttcgcta aatactggca ggcgtttcgt
     2221 cagtatcccc gtttacaggg cggcttcgtc tgggactggg tggatcagtc gctgattaaa
     2281 tatgatgaaa acggcaaccc gtggtcggct tacggcggtg attttggcga tacgccgaac
     2341 gatcgccagt tctgtatgaa cggtctggtc tttgccgacc gcacgccgca tccagcgtaa
     2401 gtttaaacaa taacctacta actaacgtag ataatttaaa ttttcaggct gacggaagca
     2461 aaacaccagc agcagttttt ccagttccgt ttatccgggc aaaccatcga agtgaccagc
     2521 gaatacctgt tccgtcatag cgataacgag ctcctgcact ggatggtggc gctggatggt
     2581 aagccgctgg caagcggtga agtgcctctg gatgtcgctc cacaaggtaa acagttgatt
     2641 gaactgcctg aactaccgca gccggagagc gccgggcaac tctggctcac agtacgcgta
     2701 gtgcaaccga acgcgaccgc atggtcagaa gccgggcaca tcagcgcatg gcagcagtgg
     2761 aggtaagttt aaacaagatc ctactaacta actctacatt gatgaatttt cagactggcg
     2821 gaaaacctca gtgtgacgct ccccgccgcg tcccacgcca tcccgcatct gaccaccagc
     2881 gaaatggatt tttgcatcga gctgggtaat aagcgttggc aatttaaccg ccagtcaggc
     2941 tttctttcac agatgtggat tggcgataaa aaacaactgc tgacgccgct gcgcgatcag
     3001 ttcacccgtg caccgctgga taacgacatt ggcgtaagtg aagcgacccg cattgaccct
     3061 aacgcctggg tcggtaagtt taaacaaagt tgtactaact aacgaagatc ttgataattt
     3121 tcagaacgct ggaaggcggc gggccattac caggccgaag cagcgttgtt gcagtgcacg
     3181 gcagatacac ttgctgatgc ggtgctgatt acgaccgctc acgcgtggca gcatcagggg
     3241 aaaaccttat ttatcagccg gaaaacctac cggattgatg gtagtggtca aatggcgatt
     3301 accgttgatg ttgaagtggc gagcgataca ccgcatccgg cgcggattgg cctgaactgc
     3361 cagctggcgc aggtagcaga gcgggtaaac tggctcggat tagggccgca agaaaactat
     3421 cccgaccgcc ttactgccgc ctgttttgac cgctgggatc tgccattgtc agacatgtag
     3481 taagtttaaa cttgatagta ctaactaaca tgtttcattt aaattttcag taccccgtac
     3541 gtcttcccga gcgaaaacgg tctgcgctgc gggacgcgcg aattgaatta tggcccacac
     3601 cagtggcgcg gcgacttcca gttcaacatc agccgctaca gtcaacagca actgatggaa
     3661 accagccatc gccatctgct gcacgcggaa gaaggcacat ggctgaatat cgacggtttc
     3721 catatgggga ttggtggcga cgactcctgg agcccgtcag tatcggcgga attccaactg
     3781 agcgccggtc gctaccatta ccaacttgtc tggtgtcaaa aataataggg gccgctgtca
     3841 tcagagtaag tttaaactga gttctactaa ctaacgagta atatttaaat tttcagcatc
     3901 tcgcgcccgt gcctctgact tctaagtcca attactcttc aacatcccta catgctcttt
     3961 ctccctgtgc tcccaccccc tatttttgtt attatcaaaa aacttctctt aatttctttg
     4021 ttttttagct tcttttaagt cacctctaac aatgaaattg tgtagattca aaaatagaat
     4081 taattcgtaa taaaaagtcg aaaaaaattg tgctccctcc ccccattaat aataattcta
     4141 tcccaaaatc tacacaatgt tctgtgtaca cttcttatgt tttttacttc tgataaattt
     4201 ttttgaaaca tcatagaaaa aaccgcacac aaaatacctt atcatatgtt acgtttcagt
     4261 ttatgaccgc aatttttatt tcttcgcacg tctgggcctc tcatgacgtc aaatcatgct
     4321 catcgtgaaa aagttttgga gtatttttgg aatttttcaa tcaagtgaaa gtttatgaaa
     4381 ttaattttcc tgcttttgct ttttggggtt tcccctattg tttgtcaaga tttcgaggac
     4441 ggcgtttttc ttgctaaaat cacaagtatt gatgagcacg atgcaagaaa gatcggaaga
     4501 aggtttgggt ttgaggctca gtggaaggtg agtagaagtt gataatttga aagtggagta
     4561 gtgtctatgg ggtttttgcc ttaaatgaca gaatacattc ccaatatacc aaacataact
     4621 gtttcctact agtcggccgt acgggccctt tcgtctcgcg cgtttcggtg atgacggtga
     4681 aaacctctga cacatgcagc tcccggagac ggtcacagct tgtctgtaag cggatgccgg
     4741 gagcagacaa gcccgtcagg gcgcgtcagc gggtgttggc gggtgtcggg gctggcttaa
     4801 ctatgcggca tcagagcaga ttgtactgag agtgcaccat atgcggtgtg aaataccgca
     4861 cagatgcgta aggagaaaat accgcatcag gcggccttaa gggcctcgtg atacgcctat
     4921 ttttataggt taatgtcatg ataataatgg tttcttagac gtcaggtggc acttttcggg
     4981 gaaatgtgcg cggaacccct atttgtttat ttttctaaat acattcaaat atgtatccgc
     5041 tcatgagaca ataaccctga taaatgcttc aataatattg aaaaaggaag agtatgagta
     5101 ttcaacattt ccgtgtcgcc cttattccct tttttgcggc attttgcctt cctgtttttg
     5161 ctcacccaga aacgctggtg aaagtaaaag atgctgaaga tcagttgggt gcacgagtgg
     5221 gttacatcga actggatctc aacagcggta agatccttga gagttttcgc cccgaagaac
     5281 gttttccaat gatgagcact tttaaagttc tgctatgtgg cgcggtatta tcccgtattg
     5341 acgccgggca agagcaactc ggtcgccgca tacactattc tcagaatgac ttggttgagt
     5401 actcaccagt cacagaaaag catcttacgg atggcatgac agtaagagaa ttatgcagtg
     5461 ctgccataac catgagtgat aacactgcgg ccaacttact tctgacaacg atcggaggac
     5521 cgaaggagct aaccgctttt ttgcacaaca tgggggatca tgtaactcgc cttgatcgtt
     5581 gggaaccgga gctgaatgaa gccataccaa acgacgagcg tgacaccacg atgcctgtag
     5641 caatggcaac aacgttgcgc aaactattaa ctggcgaact acttactcta gcttcccggc
     5701 aacaattaat agactggatg gaggcggata aagttgcagg accacttctg cgctcggccc
     5761 ttccggctgg ctggtttatt gctgataaat ctggagccgg tgagcgtggg tctcgcggta
     5821 tcattgcagc actggggcca gatggtaagc cctcccgtat cgtagttatc tacacgacgg
     5881 ggagtcaggc aactatggat gaacgaaata gacagatcgc tgagataggt gcctcactga
     5941 ttaagcattg gtaactgtca gaccaagttt actcatatat actttagatt gatttaaaac
     6001 ttcattttta atttaaaagg atctaggtga agatcctttt tgataatctc atgaccaaaa
     6061 tcccttaacg tgagttttcg ttccactgag cgtcagaccc cgtagaaaag atcaaaggat
     6121 cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt gcaaacaaaa aaaccaccgc
     6181 taccagcggt ggtttgtttg ccggatcaag agctaccaac tctttttccg aaggtaactg
     6241 gcttcagcag agcgcagata ccaaatactg tccttctagt gtagccgtag ttaggccacc
     6301 acttcaagaa ctctgtagca ccgcctacat acctcgctct gctaatcctg ttaccagtgg
     6361 ctgctgccag tggcgataag tcgtgtctta ccgggttgga ctcaagacga tagttaccgg
     6421 ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac acagcccagc ttggagcgaa
     6481 cgacctacac cgaactgaga tacctacagc gtgagcattg agaaagcgcc acgcttcccg
     6541 aagggagaaa ggcggacagg tatccggtaa gcggcagggt cggaacagga gagcgcacga
     6601 gggagcttcc agggggaaac gcctggtatc tttatagtcc tgtcgggttt cgccacctct
     6661 gacttgagcg tcgatttttg tgatgctcgt caggggggcg gagcctatgg aaaaacgcca
     6721 gcaacgcggc ctttttacgg ttcctggcct tttgctggcc ttttgctcac atgttctttc
     6781 ctgcgttatc ccctgattct gtggataacc gtattaccgc ctttgagtga gctgataccg
     6841 ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc
     6901 caatacgcaa accgcctctc cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca
     6961 ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat taatgtgagt tagctcactc
     7021 attaggcacc ccaggcttta cactttatgc ttccggctcg tatgttgtgt ggaattgtga
     7081 gcggataaca atttcacaca ggaaacagct
//
