LOCUS       sequence-53-expo        4427 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 4427)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1995
REFERENCE   2  (bases 1 to 4427)
  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: pPD30_35
FEATURES             Location/Qualifiers
     source          1..4427
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2001..2023)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     2123..2142
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(2220..2238)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        2306..2410
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2411..2479,2480..3271)
                     /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(2629..2648)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3442..4030
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3931..3950
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     4184..4201
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    4318..4339
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(4354..4359,4360..4377,4378..4384)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4392..4408
                     /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     4397..4419
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(4416..4427,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(4416..4427,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgcatgcc tgcagtgagt attttcggtt tttcactggt
       61 tctctgtaaa aagagtaatg caaagggcaa gttaacttag gtcgtaaatg tattgaattt
      121 gcttaaaatc tgaagatcta gtggtgaacc gtggaagatt atcaagagga ggctgaagat
      181 ctgtttaaga accattaatc aaactggtat tctattttca ctggttgtat gtaaacattc
      241 tatcttattc cttttatcac tgttctgcac tttcctataa aaaaagttga ccgaccgtac
      301 tctctgaatt catttttccc gatcttacca actcccgatc tatctctatc cctggttttt
      361 tcttcgtgct ccaatggaat tcttgagact tccactatct tctctggcac cctccactac
      421 gcgtaggcgt ctctcgcttc gtgtattccc gggaagccgg gttcccgtct ctcccgccgc
      481 tgccgctgcc gcacacagct ttacacctcg tagaatcccc aaagaggggc gtggcttgcg
      541 ggtgccaaca tcctcctgcc gaggaagaag caggcactca tcactcgcat catcaacctc
      601 gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac atagaacatt
      661 ttcaggagga cccttggcta gcgtcgacgg taccatggta ttgatatctg agctccgcat
      721 cggccgctgt catcagatcg ccatctcgcg cccgtgcctc tgacttctaa gtccaattac
      781 tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt ttgttattat
      841 caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct ctaacaatga
      901 aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc
      961 cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt gtacacttct
     1021 tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg cacacaaaat
     1081 accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc gcacgtctgg
     1141 gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt tttggaattt
     1201 ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg gggtttcccc
     1261 tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa gtattgatga
     1321 gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga aggtgagtag
     1381 aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa tgacagaata
     1441 cattcccaat ataccaaaca taactgttta aaattaaaca tttttctaaa ttttatatga
     1501 tttcttttaa atttgcaaaa attacttaaa tttgaattcc cgcgcaaatg agtgacttca
     1561 ttttctgcat tattgtgttt tccggctata ttaataggta tttgtttgtg tttttcttta
     1621 ttttatgatt cgaactccaa tttgtaaatt ttcgaacata tttccctaaa gaaaaaatat
     1681 gattaatctg gaaaaattgg aaaattattt ttcaaataaa aaacaaagaa aaaaatgaag
     1741 aaaaacctat tagtttggcc ataaaacgca aaaatgtcga aaatgacgtc actcatctgc
     1801 gcgggaaatc aagaataatt cggccttttt tatttttttg gaaaatcgta aaacatttag
     1861 aaaaattttt taatagttat agtgggactg tattctgtca tttagggcaa aagccagaga
     1921 cgctactcca ccgttggggg atccactagt cggccgtacg ggccctttcg tctcgcgcgt
     1981 ttcggtgatg acggtgaaaa cctctgacac atgcagctcc cggagacggt cacagcttgt
     2041 ctgtaagcgg atgccgggag cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg
     2101 tgtcggggct ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatatg
     2161 cggtgtgaaa taccgcacag atgcgtaagg agaaaatacc gcatcaggcg gccttaaggg
     2221 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     2281 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     2341 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     2401 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     2461 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     2521 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     2581 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     2641 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     2701 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     2761 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     2821 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     2881 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     2941 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     3001 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     3061 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     3121 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     3181 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     3241 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     3301 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     3361 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     3421 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     3481 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     3541 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     3601 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     3661 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     3721 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     3781 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agcattgaga
     3841 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     3901 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     3961 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     4021 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     4081 tgctcacatg ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt
     4141 tgagtgagct gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga
     4201 ggaagcggaa gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta
     4261 atgcagctgg cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa
     4321 tgtgagttag ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat
     4381 gttgtgtgga attgtgagcg gataacaatt tcacacagga aacagct
//
