LOCUS       sequence-272-exp        5731 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 5731)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 5731)
  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: L4640
FEATURES             Location/Qualifiers
     source          1..5731
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1353..1381)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     2129..2149
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(3171..3193)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     3293..3312
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(3390..3408)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3476..3580
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3581..3649,3650..4441)
                     /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(3799..3818)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4612..5200
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5101..5120
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5354..5371
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5488..5509
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5524..5529,5530..5547,5548..5554)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5562..5578
                     /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     5567..5589
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5586..5602
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5586..5602
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttgcat gcctgcaggt cgactctaga ggatccccag cttgcatgcc tgcaggtcga
       61 ggcatttgaa ttgggggtgg tggacagtaa ctgtctgtaa taataattac tcctgaccag
      121 gttgcaattc gagttttgat aagcataatt ataccttgta cattgtgggt tttgtgctgt
      181 ggacgtttta ttgtgacatc cccataagct acaagaacaa aaatgaaatg taaaagtatt
      241 gaaaacgtcg tacattttat atctgagtag tatcctttgc tttaaatgtc cataaaacta
      301 attttataat caataaaaca acgtttgtaa atcaactgag tttacaagta gagacattga
      361 gggatacttt cactatgcta aagtgataat cgaccaaata ataacttcac tttggtattt
      421 attcctgtct tataaatgtt atgtatgaat taaattcata tgcatatggc tcactctgac
      481 aatttttaat aatcttccag atcaatattg actaccgatg cgggtggtct tttgctttga
      541 attctgctga actttacacc ccgaacagca atgtgtgctt cagcctaaaa aaaagtaagt
      601 gtgttaatca gtgcccccga ttcttcattt tttgcccctc tctcccgttt cgtcggcaaa
      661 agaagagaaa ataaagataa gtctcaagat aggttggtaa tcgctaaagt ggttgtgtgg
      721 ataagagtag caaaatggca ggaagagcac tttgcgcgca cacactgtac tcattgttct
      781 ggataaaatt ctctcgttgt ttgccgtcgg atgtctgcct ctctgcattg agccggcttc
      841 ttcactatct ttagttaacc taaaatgccg tttcttttct cgtatcccca ctatcccgtt
      901 gaggttctct gctctcttcg ctccctaccg ccagcgagca actatccgtg ggggcgcctt
      961 gctcggaaga tgggggggaa gaaagaagat ttttgctatt tgcacttgag aaagagactt
     1021 ttcctgcgtc gatggttaga gaacagtgtg cagacacttt tcagctacct agaattacaa
     1081 ttggatatcc ccgcctccca atccacccac ccagggaaaa agaagggctc gccgaaaatc
     1141 aaagttatct ccaggctcgc gcatcccacc gagcggttga cttctctcca ccacttttca
     1201 ttttaaccct cggggtacgg gattggccaa aggacccaaa ggtatgtttc gaatgatact
     1261 aacataacat agaacatttt caggaggacc cttggctagc gtcgacggta ccggtagaaa
     1321 aaatgagtaa aggagaagaa cttttcactg gagttgtccc aattcttgtt gaattagatg
     1381 gtgatgttaa tgggcacaaa ttttctgtca gtggagaggg tgaaggtgat gcaacatacg
     1441 gaaaacttac ccttaaattt atttgcacta ctggaaaact acctgttcca tgggtaagtt
     1501 taaacatata tatactaact aaccctgatt atttaaattt tcagccaaca cttgtcacta
     1561 ctttcggtta tggtcttcaa tgcttcgcca gatacccaga tcatatgaaa cagcatgact
     1621 ttttcaagag tgccatgccc gaaggttatg tacaggaaag aactatattt ttcaaagatg
     1681 acgggaacta caagacacgt aagtttaaac agttcggtac taactaacca tacatattta
     1741 aattttcagg tgctgaagtc aagtttgaag gtgataccct tgttaataga atcgagttaa
     1801 aaggtattga ttttaaagaa gatggaaaca ttcttggaca caaattggaa tacaactata
     1861 actcacacaa tgtatacatc atggcagaca aacaaaagaa tggaatcaaa gttgtaagtt
     1921 taaacatgat tttactaact aactaatctg atttaaattt tcagaacttc aaaattagac
     1981 acaacattga agatggaagc gttcaactag cagaccatta tcaacaaaat actccaattg
     2041 gcgatggccc tgtcctttta ccagacaacc attacctgtc ctaccaatct gccctttcga
     2101 aagatcccaa cgaaaagaga gaccacatgg tccttcttga gtttgtaaca gctgctggga
     2161 ttacacatgg catggatgaa ctatacaaat agcattcgta gaattgtaag tttaaacttt
     2221 ctcatactaa ctaacgcgct ctatttaaat tttcagaatt ccaactgagc gccggtcgct
     2281 accattacca acttgtctgg tgtcaaaaat aataggggcc gctgtcatca gagtaagttt
     2341 aaactgagtt ctactaacta acgagtaata tttaaatttt cagcatctcg cgcccgtgcc
     2401 tctgacttct aagtccaatt actcttcaac atccctacat gctctttctc cctgtgctcc
     2461 caccccctat ttttgttatt atcaaaaaac ttctcttaat ttctttgttt tttagcttct
     2521 tttaagtcac ctctaacaat gaaattgtgt agattcaaaa atagaattaa ttcgtaataa
     2581 aaagtcgaaa aaaattgtgc tccctccccc cattaataat aattctatcc caaaatctac
     2641 acaatgttct gtgtacactt cttatgtttt ttacttctga taaatttttt tgaaacatca
     2701 tagaaaaaac cgcacacaaa ataccttatc atatgttacg tttcagttta tgaccgcaat
     2761 ttttatttct tcgcacgtct gggcctctca tgacgtcaaa tcatgctcat cgtgaaaaag
     2821 ttttggagta tttttggaat ttttcaatca agtgaaagtt tatgaaatta attttcctgc
     2881 ttttgctttt tggggtttcc cctattgttt gtcaagattt cgaggacggc gtttttcttg
     2941 ctaaaatcac aagtattgat gagcacgatg caagaaagat cggaagaagg tttgggtttg
     3001 aggctcagtg gaaggtgagt agaagttgat aatttgaaag tggagtagtg tctatggggt
     3061 ttttgcctta aatgacagaa tacattccca atataccaaa cataactgtt tcctactagt
     3121 cggccgtacg ggccctttcg tctcgcgcgt ttcggtgatg acggtgaaaa cctctgacac
     3181 atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag cagacaagcc
     3241 cgtcagggcg cgtcagcggg tgttggcggg tgtcggggct ggcttaacta tgcggcatca
     3301 gagcagattg tactgagagt gcaccatatg cggtgtgaaa taccgcacag atgcgtaagg
     3361 agaaaatacc gcatcaggcg gccttaaggg cctcgtgata cgcctatttt tataggttaa
     3421 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     3481 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     3541 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     3601 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     3661 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     3721 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     3781 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtattgacg ccgggcaaga
     3841 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     3901 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     3961 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     4021 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     4081 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgtagcaa tggcaacaac
     4141 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     4201 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     4261 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     4321 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     4381 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     4441 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     4501 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     4561 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     4621 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     4681 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     4741 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     4801 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     4861 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     4921 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     4981 actgagatac ctacagcgtg agcattgaga aagcgccacg cttcccgaag ggagaaaggc
     5041 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     5101 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     5161 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     5221 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     5281 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     5341 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcccaa tacgcaaacc
     5401 gcctctcccc gcgcgttggc cgattcatta atgcagctgg cacgacaggt ttcccgactg
     5461 gaaagcgggc agtgagcgca acgcaattaa tgtgagttag ctcactcatt aggcacccca
     5521 ggctttacac tttatgcttc cggctcgtat gttgtgtgga attgtgagcg gataacaatt
     5581 tcacacagga aacagctatg accatgatta cgccaagctg taagtttaaa catgatctta
     5641 ctaactaact attctcattt aaattttcag agcttaaaaa tggctgaaat cactcacaac
     5701 gatggatacg ctaacaactt ggaaatgaaa t
//
