LOCUS       sequence-152-exp        6859 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 6859)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1997
REFERENCE   2  (bases 1 to 6859)
  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: L3513
FEATURES             Location/Qualifiers
     source          1..6859
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2536..2564)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3312..3332
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4299..4321)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4421..4440
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4518..4536)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4604..4708
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4709..4777,4778..5569)
                     /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(4927..4946)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5740..6328
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6229..6248
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6482..6499
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6616..6637
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6652..6657,6658..6675,6676..6682)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6690..6706
                     /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     6695..6717
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     6714..6730
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     6714..6730
                     /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 gtaccggtag aaaaaatgag taaaggagaa
     2521 gaacttttca ctggagttgt cccaattctt gttgaattag atggtgatgt taatgggcac
     2581 aaattttctg tcagtggaga gggtgaaggt gatgcaacat acggaaaact tacccttaaa
     2641 tttatttgca ctactggaaa actacctgtt ccatgggtaa gtttaaacat atatatacta
     2701 actaaccctg attatttaaa ttttcagcca acacttgtca ctactttctc acacggcgta
     2761 caatgcttct cgagataccc agatcatatg aaacggcatg actttttcaa gagtgccatg
     2821 cccgaaggtt atgtacagga aagaactata tttttcaaag atgacgggaa ctacaagaca
     2881 cgtaagttta aacagttcgg tactaactaa ccatacatat ttaaattttc aggtgctgaa
     2941 gtcaagtttg aaggtgatac ccttgttaat agaatcgagt taaaaggtat tgattttaaa
     3001 gaagatggaa acattcttgg acacaaattg gaatacaact ataactcaca caatgtatac
     3061 atcatggcag acaaacaaaa gaatggaatc aaagttgtaa gtttaaacat gattttacta
     3121 actaactaat ctgatttaaa ttttcagaac ttcaaaatta gacacaacat tgaagatgga
     3181 agcgttcaac tagcagacca ttatcaacaa aatactccaa ttggcgatgg ccctgtcctt
     3241 ttaccagaca accattacct gtccacacaa tctgcccttt cgaaagatcc caacgaaaag
     3301 agagaccaca tggtccttct tgagtttgta acagctgctg ggattacaca tggcatggat
     3361 gaactataca aatagcattc gtagaattcc aactgagcgc cggtcgctac cattaccaac
     3421 ttgtctggtg tcaaaaataa taggggccgc tgtcatcaga gtaagtttaa actgagttct
     3481 actaactaac gagtaatatt taaattttca gcatctcgcg cccgtgcctc tgacttctaa
     3541 gtccaattac tcttcaacat ccctacatgc tctttctccc tgtgctccca ccccctattt
     3601 ttgttattat caaaaaactt ctcttaattt ctttgttttt tagcttcttt taagtcacct
     3661 ctaacaatga aattgtgtag attcaaaaat agaattaatt cgtaataaaa agtcgaaaaa
     3721 aattgtgctc cctcccccca ttaataataa ttctatccca aaatctacac aatgttctgt
     3781 gtacacttct tatgtttttt acttctgata aatttttttg aaacatcata gaaaaaaccg
     3841 cacacaaaat accttatcat atgttacgtt tcagtttatg accgcaattt ttatttcttc
     3901 gcacgtctgg gcctctcatg acgtcaaatc atgctcatcg tgaaaaagtt ttggagtatt
     3961 tttggaattt ttcaatcaag tgaaagttta tgaaattaat tttcctgctt ttgctttttg
     4021 gggtttcccc tattgtttgt caagatttcg aggacggcgt ttttcttgct aaaatcacaa
     4081 gtattgatga gcacgatgca agaaagatcg gaagaaggtt tgggtttgag gctcagtgga
     4141 aggtgagtag aagttgataa tttgaaagtg gagtagtgtc tatggggttt ttgccttaaa
     4201 tgacagaata cattcccaat ataccaaaca taactgtttc ctactagtcg gccgtacggg
     4261 ccctttcgtc tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg
     4321 gagacggtca cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg
     4381 tcagcgggtg ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta
     4441 ctgagagtgc accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc
     4501 atcaggcggc cttaagggcc tcgtgatacg cctattttta taggttaatg tcatgataat
     4561 aatggtttct tagacgtcag gtggcacttt tcggggaaat gtgcgcggaa cccctatttg
     4621 tttatttttc taaatacatt caaatatgta tccgctcatg agacaataac cctgataaat
     4681 gcttcaataa tattgaaaaa ggaagagtat gagtattcaa catttccgtg tcgcccttat
     4741 tccctttttt gcggcatttt gccttcctgt ttttgctcac ccagaaacgc tggtgaaagt
     4801 aaaagatgct gaagatcagt tgggtgcacg agtgggttac atcgaactgg atctcaacag
     4861 cggtaagatc cttgagagtt ttcgccccga agaacgtttt ccaatgatga gcacttttaa
     4921 agttctgcta tgtggcgcgg tattatcccg tattgacgcc gggcaagagc aactcggtcg
     4981 ccgcatacac tattctcaga atgacttggt tgagtactca ccagtcacag aaaagcatct
     5041 tacggatggc atgacagtaa gagaattatg cagtgctgcc ataaccatga gtgataacac
     5101 tgcggccaac ttacttctga caacgatcgg aggaccgaag gagctaaccg cttttttgca
     5161 caacatgggg gatcatgtaa ctcgccttga tcgttgggaa ccggagctga atgaagccat
     5221 accaaacgac gagcgtgaca ccacgatgcc tgtagcaatg gcaacaacgt tgcgcaaact
     5281 attaactggc gaactactta ctctagcttc ccggcaacaa ttaatagact ggatggaggc
     5341 ggataaagtt gcaggaccac ttctgcgctc ggcccttccg gctggctggt ttattgctga
     5401 taaatctgga gccggtgagc gtgggtctcg cggtatcatt gcagcactgg ggccagatgg
     5461 taagccctcc cgtatcgtag ttatctacac gacggggagt caggcaacta tggatgaacg
     5521 aaatagacag atcgctgaga taggtgcctc actgattaag cattggtaac tgtcagacca
     5581 agtttactca tatatacttt agattgattt aaaacttcat ttttaattta aaaggatcta
     5641 ggtgaagatc ctttttgata atctcatgac caaaatccct taacgtgagt tttcgttcca
     5701 ctgagcgtca gaccccgtag aaaagatcaa aggatcttct tgagatcctt tttttctgcg
     5761 cgtaatctgc tgcttgcaaa caaaaaaacc accgctacca gcggtggttt gtttgccgga
     5821 tcaagagcta ccaactcttt ttccgaaggt aactggcttc agcagagcgc agataccaaa
     5881 tactgtcctt ctagtgtagc cgtagttagg ccaccacttc aagaactctg tagcaccgcc
     5941 tacatacctc gctctgctaa tcctgttacc agtggctgct gccagtggcg ataagtcgtg
     6001 tcttaccggg ttggactcaa gacgatagtt accggataag gcgcagcggt cgggctgaac
     6061 ggggggttcg tgcacacagc ccagcttgga gcgaacgacc tacaccgaac tgagatacct
     6121 acagcgtgag cattgagaaa gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc
     6181 ggtaagcggc agggtcggaa caggagagcg cacgagggag cttccagggg gaaacgcctg
     6241 gtatctttat agtcctgtcg ggtttcgcca cctctgactt gagcgtcgat ttttgtgatg
     6301 ctcgtcaggg gggcggagcc tatggaaaaa cgccagcaac gcggcctttt tacggttcct
     6361 ggccttttgc tggccttttg ctcacatgtt ctttcctgcg ttatcccctg attctgtgga
     6421 taaccgtatt accgcctttg agtgagctga taccgctcgc cgcagccgaa cgaccgagcg
     6481 cagcgagtca gtgagcgagg aagcggaaga gcgcccaata cgcaaaccgc ctctccccgc
     6541 gcgttggccg attcattaat gcagctggca cgacaggttt cccgactgga aagcgggcag
     6601 tgagcgcaac gcaattaatg tgagttagct cactcattag gcaccccagg ctttacactt
     6661 tatgcttccg gctcgtatgt tgtgtggaat tgtgagcgga taacaatttc acacaggaaa
     6721 cagctatgac catgattacg ccaagctgta agtttaaaca tgatcttact aactaactat
     6781 tctcatttaa attttcagag cttaaaaatg gctgaaatca ctcacaacga tggatacgct
     6841 aacaacttgg aaatgaaat
//
