LOCUS       sequence-321-exp        6420 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 6420)
  TITLE     Fire Lab C. Elegans Vector Kit 2005
REFERENCE   2  (bases 1 to 6420)
  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: pPD158.114
FEATURES             Location/Qualifiers
     source          1..6420
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3860..3882)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3982..4001
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4079..4097)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4165..4269
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4270..4338,4339..5130)
                     /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(4488..4507)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5301..5889
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5790..5809
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6043..6060
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6177..6198
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6213..6218,6219..6236,6237..6243)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6251..6267
                     /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     6256..6278
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6275..6291
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6275..6291
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgggc tgcaggtcgg ctataataag ttcttgaata aaataatttt cccgacaaaa
       61 catgagtatt tctttcgaaa ataaaagtgc aggctaatta gagattattc tgtaattaac
      121 tgcataattt gtcacgtgcc atagttttac attccactac gtcatagttc ttaaaatact
      181 aatctcctga aaatagaagt aggtgaagaa agtttaatta tcagttctaa aatgacaatt
      241 gatctttgga atatgttctg aaactaccga tcattgaaca gatgctattt gaatgatata
      301 gaattgtata tttgcaattt ctgaaacgcg ttcttaaagg cacacagatt aattcaaaag
      361 ggtctggccg caaaaaggtt tatggtggcc gattttgagt tttgtgtgtg attgcttttt
      421 cacaatcagt gttttcagga ttatgtgatg aactagatct tcaagtttcg ttacatttca
      481 tatgttttcg gaactcacga agtacatatt gggtattgtg ctcaaaaaat tcagcaatca
      541 gcttcgctcc gctgacttta gaacccaaaa aaatagtatg gccaaactga ctgtgttacg
      601 atcatttcaa tttttcaata catatttaag atttctaaga gtaagaaggt caaaaactgt
      661 tctggaatac atatatattt ttcaggttac aattagtcaa aaagtgcact gaaatatacg
      721 ttttaatttc acgaataacc caattagttc aatgtatttt tggtcaacca acgttaaagt
      781 ttggcttcca accaattatc atttctgatc aaccacaatg ttttttcttt atctgcaagt
      841 taatttttta tttttatcca gatgtttggc atatttttca attcttcact agcgcccact
      901 tcttgcactt ccggcgccct gaatctaatg catctgttgc aagaattgaa agaccaatca
      961 acacattgtt ttcttcacga gatactgaag aaaatgaata aaaacagaga aaaagagcca
     1021 tgtgattagt gacaactgtt gctaacagat accatagctt ggacttggta cgtgatggca
     1081 acgtatgggt caacaaaaat gattgcagag ggggtgcaaa acagtcaagt cgagaaaata
     1141 tgaaaaacag aaaacaaaga acagaaaaat gggtttgaga gtcagtataa tttataaaag
     1201 aaaaattgta catagaaatt aaccattttt gtagaagaag ttatttttca agcatcgtta
     1261 aaaattattc aaagcacctt atttcatatt taattttaaa catggttaaa tgaacaacac
     1321 ggtgcgcaat caggaaaact tgaaatctga aactgttgtt gtgatcttct tcgcaactgt
     1381 tcagatagca ctagtgtaat gttaagagtg cgcgaatata atggaatata atggatcaca
     1441 cctcctgcca tcaggtaaac gtctctgtta tcacatattt ccaactatta aatttttacc
     1501 ttttacagtt ttacattttt ttgaaaaaag taactttttg tcttcaaaat ccctgacgaa
     1561 aatatcaaat attttaatcg agactgcaga ggaaccgatt gatgatttgg aaaatccagc
     1621 tttacctgtg taagaactga aaagtttcat aaccctaggg tattcccagt tacattcccc
     1681 actggctaac aatagcaccc agtttttcat cacctttctt caaatttctc ggcgatttgt
     1741 taaaaacaaa atttgtgtcc cttctctgat atctctatgt ctctaaacac aagttcatcg
     1801 gaaaacgaag gagggtaggt gttggttggg ctcccgaagt gaaaatagaa gagcaagaat
     1861 agaatattag agagagagtg cagagagggc gggatagctc ccgggattcc gttttcttct
     1921 tctttatctt caacgatgat gtgtgtgcgt gttgtataga ttctgttgct cccccacaac
     1981 tcgctccgaa ggctcaatac aattcaattg atattggagg agagcctacc ggagtgggag
     2041 gataagaaga aacataagaa gaagaagaag aagaagcatg cttctggttt ttgatgctat
     2101 gaaaacggca caaaaagatg attgaggtcc cttttcaata ccttctctca tctttcaaat
     2161 cccattgaaa cctaaaactt ctcaccacgc tttaccattg ttctccaaaa acttatagca
     2221 atgtctataa cttttttatc tctgaaaagc agtgttccat ttttcttttt cctattttat
     2281 ttcaattgtt tctcacattt cgtttggatt ctttgcttgt caaccagctt cttcttccac
     2341 ttttaccgtc taattttcag ggcagggagc catcaaaccc acgaccacta gatccatcta
     2401 gaggatcccc gggattggcc aaaggaccca aaggtatgtt tcgaatgata ctaacataac
     2461 atagaacatt ttcaggagga cccttggagg gtaccggtcg ccaccatgag gtcttccaag
     2521 aatgttatca aggagttcat gaggtttaag gttcgcatgg aaggaacggt caatgggcac
     2581 gagtttgaaa tagaaggcga aggagagggg aggccatacg aaggccacaa taccgtaaag
     2641 cttaaggtaa ccaagggggg acctttgcca tttgcttggg atattttgtc accacaattt
     2701 cagtatggaa gcaaggtata tgtcaagcac cctgccgaca taccagacta taaaaagctg
     2761 tcatttcctg aaggatttgt aagtttaaac ggggtgttac taactaacaa ataacattta
     2821 aattttcaga aatgggaaag ggtcatgaac tttgaagacg gtggcgtcgc tactgtcacc
     2881 caagattcca gcttgcaaga tggctgtttc atctacaagg tcaagttcat tggcgtgaac
     2941 tttccttccg atggacctgt tatgcaaaag aagacaatgg gctgggaagc cagcactgag
     3001 cgtttgtatc ctcgtgatgg cgtgttgaaa ggagagattc ataaggctct gaagctgaaa
     3061 gacgggtaag tttaaactat atcttactaa ctaactgtag agatttaaat tttcagtggt
     3121 cattacctag ttgaattcaa aagtatttac atggcaaaga agcctgtgca gctaccaggg
     3181 tactactatg ttgataccaa actggatatc acaagccaca acgaagacta tacaatcgtt
     3241 gagcagtatg aaagaaccga gggacgccac catctgttcc tttagcggcc gcgactctag
     3301 aattccaact gagcgccggt cgctaccatt accaacttgt ctggtgtcaa aaataatagg
     3361 cctactagcc ggccatacaa gtaatccgga tgatcgacgc caacgtcgtt gaattttcaa
     3421 attttaaata ctgaatattt gttttttttc ctattattta tttattctct ttgtgttttt
     3481 tttcttgctt tctaaaaaat taattcaatc caaatctaaa catttttttt tctctttccg
     3541 tctcccaatt cgtattccgc tcctctcatc tgaacacaat gtgcaagttt atttatcttc
     3601 tcgctttcat ttcattagga cgtgggggga attggtggaa gggggaaaca cacaaaagga
     3661 tgatggaaat gaaataagga cacacaatat gcaacaacat tcaattcaga aatatggagg
     3721 aaggtttaaa agaaaacata aaaatatata gaggaggaag gaaaactagt aaaaaataag
     3781 caaagaaatt aggcgaacga tgagaattgt cctcgcttgg gccctttcgt ctcgcgcgtt
     3841 tcggtgatga cggtgaaaac ctctgacaca tgcagctccc ggagacggtc acagcttgtc
     3901 tgtaagcgga tgccgggagc agacaagccc gtcagggcgc gtcagcgggt gttggcgggt
     3961 gtcggggctg gcttaactat gcggcatcag agcagattgt actgagagtg caccatatgc
     4021 ggtgtgaaat accgcacaga tgcgtaagga gaaaataccg catcaggcgg ccttaagggc
     4081 ctcgtgatac gcctattttt ataggttaat gtcatgataa taatggtttc ttagacgtca
     4141 ggtggcactt ttcggggaaa tgtgcgcgga acccctattt gtttattttt ctaaatacat
     4201 tcaaatatgt atccgctcat gagacaataa ccctgataaa tgcttcaata atattgaaaa
     4261 aggaagagta tgagtattca acatttccgt gtcgccctta ttcccttttt tgcggcattt
     4321 tgccttcctg tttttgctca cccagaaacg ctggtgaaag taaaagatgc tgaagatcag
     4381 ttgggtgcac gagtgggtta catcgaactg gatctcaaca gcggtaagat ccttgagagt
     4441 tttcgccccg aagaacgttt tccaatgatg agcactttta aagttctgct atgtggcgcg
     4501 gtattatccc gtattgacgc cgggcaagag caactcggtc gccgcataca ctattctcag
     4561 aatgacttgg ttgagtactc accagtcaca gaaaagcatc ttacggatgg catgacagta
     4621 agagaattat gcagtgctgc cataaccatg agtgataaca ctgcggccaa cttacttctg
     4681 acaacgatcg gaggaccgaa ggagctaacc gcttttttgc acaacatggg ggatcatgta
     4741 actcgccttg atcgttggga accggagctg aatgaagcca taccaaacga cgagcgtgac
     4801 accacgatgc ctgtagcaat ggcaacaacg ttgcgcaaac tattaactgg cgaactactt
     4861 actctagctt cccggcaaca attaatagac tggatggagg cggataaagt tgcaggacca
     4921 cttctgcgct cggcccttcc ggctggctgg tttattgctg ataaatctgg agccggtgag
     4981 cgtgggtctc gcggtatcat tgcagcactg gggccagatg gtaagccctc ccgtatcgta
     5041 gttatctaca cgacggggag tcaggcaact atggatgaac gaaatagaca gatcgctgag
     5101 ataggtgcct cactgattaa gcattggtaa ctgtcagacc aagtttactc atatatactt
     5161 tagattgatt taaaacttca tttttaattt aaaaggatct aggtgaagat cctttttgat
     5221 aatctcatga ccaaaatccc ttaacgtgag ttttcgttcc actgagcgtc agaccccgta
     5281 gaaaagatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     5341 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     5401 tttccgaagg taactggctt cagcagagcg cagataccaa atactgtcct tctagtgtag
     5461 ccgtagttag gccaccactt caagaactct gtagcaccgc ctacatacct cgctctgcta
     5521 atcctgttac cagtggctgc tgccagtggc gataagtcgt gtcttaccgg gttggactca
     5581 agacgatagt taccggataa ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag
     5641 cccagcttgg agcgaacgac ctacaccgaa ctgagatacc tacagcgtga gcattgagaa
     5701 agcgccacgc ttcccgaagg gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga
     5761 acaggagagc gcacgaggga gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc
     5821 gggtttcgcc acctctgact tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc
     5881 ctatggaaaa acgccagcaa cgcggccttt ttacggttcc tggccttttg ctggcctttt
     5941 gctcacatgt tctttcctgc gttatcccct gattctgtgg ataaccgtat taccgccttt
     6001 gagtgagctg ataccgctcg ccgcagccga acgaccgagc gcagcgagtc agtgagcgag
     6061 gaagcggaag agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa
     6121 tgcagctggc acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat
     6181 gtgagttagc tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg
     6241 ttgtgtggaa ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac
     6301 gccaagctgt aagtttaaac atgatcttac taactaacta ttctcattta aattttcaga
     6361 gcttaaaaat ggctgaaatc actcacaacg atggatacgc taacaacttg gaaatgaaat
//
