LOCUS       sequence-250-exp        6843 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 6843)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6843)
  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: L4066
FEATURES             Location/Qualifiers
     source          1..6843
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2520..2548)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3296..3316
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4283..4305)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4405..4424
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4502..4520)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4588..4692
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4693..4761,4762..5553)
                     /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(4911..4930)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5724..6312
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6213..6232
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6466..6483
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6600..6621
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6636..6641,6642..6659,6660..6666)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6674..6690
                     /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     6679..6701
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6698..6714
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6698..6714
                     /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 gaccgcggtt ttggcgcgga tcctatcgat tcgcggccgc tgtacacccg ggtgccctat
     2461 agtgagtcgt attggtaccg gtagaaaaaa tgagtaaagg agaagaactt ttcactggag
     2521 ttgtcccaat tcttgttgaa ttagatggtg atgttaatgg gcacaaattt tctgtcagtg
     2581 gagagggtga aggtgatgca acatacggaa aacttaccct taaatttatt tgcactactg
     2641 gaaaactacc tgttccatgg gtaagtttaa acatatatat actaactaac cctgattatt
     2701 taaattttca gccaacactt gtcactactc tcacttgggg cgtacaatgc ttctcgagat
     2761 acccagatca tatgaaacag catgactttt tcaagagtgc catgcccgaa ggttatgtac
     2821 aggaaagaac tatatttttc aaagatgacg ggaactacaa gacacgtaag tttaaacagt
     2881 tcggtactaa ctaaccatac atatttaaat tttcaggtgc tgaagtcaag tttgaaggtg
     2941 atacccttgt taatagaatc gagttaaaag gtattgattt taaagaagat ggaaacattc
     3001 ttggacacaa attggaatac aacttcaact cacacaacgt atacatcatg gcagacaaac
     3061 aaaagaatgg aatcaaagtt gtaagtttaa acatgatttt actaactaac taatctgatt
     3121 taaattttca gaacttcaaa attagacaca acattgaaga tggaagcgtt caactagcag
     3181 accattatca acaaaatact ccaattggcg atggccctgt ccttttacca gacaaccatt
     3241 acctgtccac acaatctgcc ctttcgaaag atcccaacga aaagagagac cacatggtcc
     3301 ttcttgagtt tgtaacagct gctgggatta cacatggcat ggatgaacta tacaaatagc
     3361 attcgtagaa ttccaactga gcgccggtcg ctaccattac caacttgtct ggtgtcaaaa
     3421 ataatagggg ccgctgtcat cagagtaagt ttaaactgag ttctactaac taacgagtaa
     3481 tatttaaatt ttcagcatct cgcgcccgtg cctctgactt ctaagtccaa ttactcttca
     3541 acatccctac atgctctttc tccctgtgct cccaccccct atttttgtta ttatcaaaaa
     3601 acttctctta atttctttgt tttttagctt cttttaagtc acctctaaca atgaaattgt
     3661 gtagattcaa aaatagaatt aattcgtaat aaaaagtcga aaaaaattgt gctccctccc
     3721 cccattaata ataattctat cccaaaatct acacaatgtt ctgtgtacac ttcttatgtt
     3781 ttttacttct gataaatttt tttgaaacat catagaaaaa accgcacaca aaatacctta
     3841 tcatatgtta cgtttcagtt tatgaccgca atttttattt cttcgcacgt ctgggcctct
     3901 catgacgtca aatcatgctc atcgtgaaaa agttttggag tatttttgga atttttcaat
     3961 caagtgaaag tttatgaaat taattttcct gcttttgctt tttggggttt cccctattgt
     4021 ttgtcaagat ttcgaggacg gcgtttttct tgctaaaatc acaagtattg atgagcacga
     4081 tgcaagaaag atcggaagaa ggtttgggtt tgaggctcag tggaaggtga gtagaagttg
     4141 ataatttgaa agtggagtag tgtctatggg gtttttgcct taaatgacag aatacattcc
     4201 caatatacca aacataactg tttcctacta gtcggccgta cgggcccttt cgtctcgcgc
     4261 gtttcggtga tgacggtgaa aacctctgac acatgcagct cccggagacg gtcacagctt
     4321 gtctgtaagc ggatgccggg agcagacaag cccgtcaggg cgcgtcagcg ggtgttggcg
     4381 ggtgtcgggg ctggcttaac tatgcggcat cagagcagat tgtactgaga gtgcaccata
     4441 tgcggtgtga aataccgcac agatgcgtaa ggagaaaata ccgcatcagg cggccttaag
     4501 ggcctcgtga tacgcctatt tttataggtt aatgtcatga taataatggt ttcttagacg
     4561 tcaggtggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     4621 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     4681 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     4741 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     4801 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     4861 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     4921 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     4981 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     5041 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     5101 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     5161 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     5221 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     5281 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     5341 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     5401 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     5461 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     5521 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     5581 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     5641 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     5701 gtagaaaaga tcaaaggatc ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg
     5761 caaacaaaaa aaccaccgct accagcggtg gtttgtttgc cggatcaaga gctaccaact
     5821 ctttttccga aggtaactgg cttcagcaga gcgcagatac caaatactgt ccttctagtg
     5881 tagccgtagt taggccacca cttcaagaac tctgtagcac cgcctacata cctcgctctg
     5941 ctaatcctgt taccagtggc tgctgccagt ggcgataagt cgtgtcttac cgggttggac
     6001 tcaagacgat agttaccgga taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca
     6061 cagcccagct tggagcgaac gacctacacc gaactgagat acctacagcg tgagcattga
     6121 gaaagcgcca cgcttcccga agggagaaag gcggacaggt atccggtaag cggcagggtc
     6181 ggaacaggag agcgcacgag ggagcttcca gggggaaacg cctggtatct ttatagtcct
     6241 gtcgggtttc gccacctctg acttgagcgt cgatttttgt gatgctcgtc aggggggcgg
     6301 agcctatgga aaaacgccag caacgcggcc tttttacggt tcctggcctt ttgctggcct
     6361 tttgctcaca tgttctttcc tgcgttatcc cctgattctg tggataaccg tattaccgcc
     6421 tttgagtgag ctgataccgc tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc
     6481 gaggaagcgg aagagcgccc aatacgcaaa ccgcctctcc ccgcgcgttg gccgattcat
     6541 taatgcagct ggcacgacag gtttcccgac tggaaagcgg gcagtgagcg caacgcaatt
     6601 aatgtgagtt agctcactca ttaggcaccc caggctttac actttatgct tccggctcgt
     6661 atgttgtgtg gaattgtgag cggataacaa tttcacacag gaaacagcta tgaccatgat
     6721 tacgccaagc tgtaagttta aacatgatct tactaactaa ctattctcat ttaaattttc
     6781 agagcttaaa aatggctgaa atcactcaca acgatggata cgctaacaac ttggaaatga
     6841 aat
//
