LOCUS       sequence-73-expo        7335 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 7335)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 7335)
  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: pPD88_75
FEATURES             Location/Qualifiers
     source          1..7335
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             386..406
                     /codon_start=1
                     /product="nuclear localization signal of SV40 (simian virus
                     40) large T antigen"
                     /label=SV40 NLS
                     /translation="PKKKRKV"
     primer_bind     complement(496..518)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(4909..4931)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     5031..5050
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(5128..5146)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5214..5318
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(5319..5387,5388..6179)
                     /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(5537..5556)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6350..6938
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6839..6858
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     7092..7109
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    7226..7247
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(7262..7267,7268..7285,7286..7292)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7300..7316
                     /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     7305..7327
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(7324..7335,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(7324..7335,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagagga tccccaattc
       61 ttgagacttc cactatcttc tctggcaccc tccactacgc gtaggcgtct ctcgcttcgt
      121 gtattcccgg gaagccgggt tcccgtctct cccgccgctg ccgctgccgc acacagcttt
      181 acacctcgta gaatccccaa agaggggcgt ggcttgcggg tgccaacatc ctcctgccga
      241 ggaagaagca ggcactcatc actcgcatca tcaacctcgg gattggccaa aggacccaaa
      301 ggtatgtttc gaatgatact aacataacat agaacatttt caggaggacc cttggagggt
      361 accgagctca gaaaaaatga ctgctccaaa gaagaagcgt aaggtaccgg tgggtgaaga
      421 ccagaaacag cacctcgaac tgagccgcga tattgcccag cgtttcaacg cgctgtatgg
      481 cgagatcgat cccgtcgttt tacaacgtcg tgactgggaa aaccctggcg ttacccaact
      541 taatcgcctt gcagcacatc cccctttcgc cagctggcgt aatagcgaag aggcccgcac
      601 cgatcgccct tcccaacagt tgcgaaggta agtttaaaca gatccatact aactaacttg
      661 ttctgacata attttcagct tgaatggcga atggcgcttt gcctggtttc cggcaccaga
      721 agcggtgccg gaaagctggc tggagtgcga tcttcctgag gccgatactg tcgtcgtccc
      781 ctcaaactgg cagatgcacg gttacgatgc gcccatctac accaacgtga cctatcccat
      841 tacggtcaat ccgccgtttg ttcccacgga gaatccgacg ggttgttact cgctcacatt
      901 taatgttgat gaaagctggc tacaggaagg ccagacgcga attatttttg atggcgttaa
      961 ctcggcgttt catctgtggt gcaacgggcg ctgggtcggt tacggccagg taagtttaat
     1021 taagttgata ctaactaaca agatctgatt aattttcagg acagtcgttt gccgtctgaa
     1081 tttgacctga gcgcattttt acgcgccgga gaaaaccgcc tcgcggtgat ggtgctgcgc
     1141 tggagtgacg gcagttatct ggaagatcag gatatgtggc ggatgagcgg cattttccgt
     1201 gacgtctcgt tgctgcataa accgactaca caaatcagcg atttccatgt tgccactcgc
     1261 tttaatgatg atttctcccg cgctgtactg gaggctgaag tccaggtaag tttaaacagg
     1321 atcttactaa ctaacatgct aacactgaat tttcagatgt gcggcgagtt gcgtgactac
     1381 ctacgggtaa cagtttcttt atggcagggt gaaacgcagg tcgccagcgg caccgcgcct
     1441 ttcggcggtg aaattatcga tgagcgtggt ggttatgccg atcgcgtcac actacgtctg
     1501 aacgtcgaaa acccgaaact gtggagcgcc gaaatcccga atctctatcg tgcggtggtt
     1561 gaactgcaca ccgccgacgg cacgctgatt gaagcagaag cctgcgatgt cggtttccgc
     1621 gaggtgcgga ttgaaaatgg tctgctgctg ctgaacggca agccgttgct gattcgaggc
     1681 gttaaccgtc acgagcatca tcctctgcat ggtcaggtca tggatgagca gacgatggtg
     1741 caggatgtaa gtttaaacta ttcgttacta actaacttta aacatttaaa ttttcagatc
     1801 ctgctgatga agcagaacaa ctttaacgcc gtgcgctgtt cgcattatcc gaaccatccg
     1861 ctgtggtaca cgctgtgcga ccgctacggc ctgtatgtgg tggatgaagc caatattgaa
     1921 acccacggca tggtgccaat gaatcgtctg accgatgatc cgcgctggct accggcgatg
     1981 agcgaacgcg taacgcgaat ggtgcagcgc gatcgtaatc acccgagtgt gatcatctgg
     2041 tcgctgggga atggtaagtt taaacagttg aatactaact aacggagatc tttgaaattt
     2101 tcagaatcag gccacggcgc taatcacgac gcgctgtatc gctggatcaa atctgtcgat
     2161 ccttcccgcc cggtgcagta tgaaggcggc ggagccgaca ccacggccac cgatattatt
     2221 tgcccgatgt acgcgcgcgt ggatgaagac cagcccttcc cggctgtgcc gaaatggtcc
     2281 atcaaaaaat ggctttcgct acctggagag acgcgcccgc tgattctttg cgaggtaagt
     2341 ttaaacagaa ctctactaac taacacatta gatcctaatt ttcagtacgc tcacgcgatg
     2401 ggcaacagtc ttggcggttt cgctaaatac tggcaggcgt ttcgtcagta tccccgttta
     2461 cagggcggct tcgtctggga ctgggtggat cagtcgctga ttaaatatga tgaaaacggc
     2521 aacccgtggt cggcttacgg cggtgatttt ggcgatacgc cgaacgatcg ccagttctgt
     2581 atgaacggtc tggtctttgc cgaccgcacg ccgcatccag cgtaagttta aacaataacc
     2641 tactaactaa cgtagataat ttaaattttc aggctgacgg aagcaaaaca ccagcagcag
     2701 tttttccagt tccgtttatc cgggcaaacc atcgaagtga ccagcgaata cctgttccgt
     2761 catagcgata acgagctcct gcactggatg gtggcgctgg atggtaagcc gctggcaagc
     2821 ggtgaagtgc ctctggatgt cgctccacaa ggtaaacagt tgattgaact gcctgaacta
     2881 ccgcagccgg agagcgccgg gcaactctgg ctcacagtac gcgtagtgca accgaacgcg
     2941 accgcatggt cagaagccgg gcacatcagc gcatggcagc agtggaggta agtttaaaca
     3001 agatcctact aactaactct acattgatga attttcagac tggcggaaaa cctcagtgtg
     3061 acgctccccg ccgcgtccca cgccatcccg catctgacca ccagcgaaat ggatttttgc
     3121 atcgagctgg gtaataagcg ttggcaattt aaccgccagt caggctttct ttcacagatg
     3181 tggattggcg ataaaaaaca actgctgacg ccgctgcgcg atcagttcac ccgtgcaccg
     3241 ctggataacg acattggcgt aagtgaagcg acccgcattg accctaacgc ctgggtcggt
     3301 aagtttaaac aaagttgtac taactaacga agatcttgat aattttcaga acgctggaag
     3361 gcggcgggcc attaccaggc cgaagcagcg ttgttgcagt gcacggcaga tacacttgct
     3421 gatgcggtgc tgattacgac cgctcacgcg tggcagcatc aggggaaaac cttatttatc
     3481 agccggaaaa cctaccggat tgatggtagt ggtcaaatgg cgattaccgt tgatgttgaa
     3541 gtggcgagcg atacaccgca tccggcgcgg attggcctga actgccagct ggcgcaggta
     3601 gcagagcggg taaactggct cggattaggg ccgcaagaaa actatcccga ccgccttact
     3661 gccgcctgtt ttgaccgctg ggatctgcca ttgtcagaca tgtagtaagt ttaaacttga
     3721 tagtactaac taacatgttt catttaaatt ttcagtaccc cgtacgtctt cccgagcgaa
     3781 aacggtctgc gctgcgggac gcgcgaattg aattatggcc cacaccagtg gcgcggcgac
     3841 ttccagttca acatcagccg ctacagtcaa cagcaactga tggaaaccag ccatcgccat
     3901 ctgctgcacg cggaagaagg cacatggctg aatatcgacg gtttccatat ggggattggt
     3961 ggcgacgact cctggagccc gtcagtatcg gcggaattcc aactgagcgc cggtcgctac
     4021 cattaccaac ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa
     4081 actgagttct actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc
     4141 tgacttctaa gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca
     4201 ccccctattt ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt
     4261 taagtcacct ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa
     4321 agtcgaaaaa aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac
     4381 aatgttctgt gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata
     4441 gaaaaaaccg cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt
     4501 ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt
     4561 ttggagtatt tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt
     4621 ttgctttttg gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct
     4681 aaaatcacaa gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag
     4741 gctcagtgga aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt
     4801 ttgccttaaa tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg
     4861 gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat
     4921 gcagctcccg gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg
     4981 tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga
     5041 gcagattgta ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag
     5101 aaaataccgc atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg
     5161 tcatgataat aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa
     5221 cccctatttg tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac
     5281 cctgataaat gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg
     5341 tcgcccttat tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc
     5401 tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg
     5461 atctcaacag cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga
     5521 gcacttttaa agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc
     5581 aactcggtcg ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag
     5641 aaaagcatct tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga
     5701 gtgataacac tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg
     5761 cttttttgca caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga
     5821 atgaagccat accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt
     5881 tgcgcaaact attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact
     5941 ggatggaggc ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt
     6001 ttattgctga taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg
     6061 ggccagatgg taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta
     6121 tggatgaacg aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac
     6181 tgtcagacca agtttactca tatatacttt agattgattt aaaacttcat ttttaattta
     6241 aaaggatcta ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt
     6301 tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt
     6361 tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt
     6421 gtttgccgga tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc
     6481 agataccaaa tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg
     6541 tagcaccgcc tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg
     6601 ataagtcgtg tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt
     6661 cgggctgaac ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac
     6721 tgagatacct acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg
     6781 acaggtatcc ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg
     6841 gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat
     6901 ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt
     6961 tacggttcct ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg
     7021 attctgtgga taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa
     7081 cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc
     7141 ctctccccgc gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga
     7201 aagcgggcag tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg
     7261 ctttacactt tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc
     7321 acacaggaaa cagct
//
