LOCUS       sequence-293-exp        6755 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 6755)
  AUTHORS   Fire A et al
  TITLE     Fire Lab C. Elegans Vector Kit 1999
REFERENCE   2  (bases 1 to 6755)
  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: L4817
FEATURES             Location/Qualifiers
     source          1..6755
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(2511..2539)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     3287..3307
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     primer_bind     complement(4329..4351)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     4451..4470
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     complement(4548..4566)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        4634..4738
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(4739..4807,4808..5599)
                     /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(4957..4976)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      5770..6358
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6259..6278
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     6512..6529
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6646..6667
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(6682..6687,6688..6705,6706..6712)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    6720..6736
                     /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     6725..6747
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     join(6744..6755,1..5)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     join(6744..6755,1..5)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 atgaccatga ttacgccaag cttgggctgc aggtcggcta taataagttc ttgaataaaa
       61 taattttccc gacaaaacat gagtatttct ttcgaaaata aaagtgcagg ctaattagag
      121 attattctgt aattaactgc ataatttgtc acgtgccata gttttacatt ccactacgtc
      181 atagttctta aaatactaat ctcctgaaaa tagaagtagg tgaagaaagt ttaattatca
      241 gttctaaaat gacaattgat ctttggaata tgttctgaaa ctaccgatca ttgaacagat
      301 gctatttgaa tgatatagaa ttgtatattt gcaatttctg aaacgcgttc ttaaaggcac
      361 acagattaat tcaaaagggt ctggccgcaa aaaggtttat ggtggccgat tttgagtttt
      421 gtgtgtgatt gctttttcac aatcagtgtt ttcaggatta tgtgatgaac tagatcttca
      481 agtttcgtta catttcatat gttttcggaa ctcacgaagt acatattggg tattgtgctc
      541 aaaaaattca gcaatcagct tcgctccgct gactttagaa cccaaaaaaa tagtatggcc
      601 aaactgactg tgttacgatc atttcaattt ttcaatacat atttaagatt tctaagagta
      661 agaaggtcaa aaactgttct ggaatacata tatatttttc aggttacaat tagtcaaaaa
      721 gtgcactgaa atatacgttt taatttcacg aataacccaa ttagttcaat gtatttttgg
      781 tcaaccaacg ttaaagtttg gcttccaacc aattatcatt tctgatcaac cacaatgttt
      841 tttctttatc tgcaagttaa ttttttattt ttatccagat gtttggcata tttttcaatt
      901 cttcactagc gcccacttct tgcacttccg gcgccctgaa tctaatgcat ctgttgcaag
      961 aattgaaaga ccaatcaaca cattgttttc ttcacgagat actgaagaaa atgaataaaa
     1021 acagagaaaa agagccatgt gattagtgac aactgttgct aacagatacc atagcttgga
     1081 cttggtacgt gatggcaacg tatgggtcaa caaaaatgat tgcagagggg gtgcaaaaca
     1141 gtcaagtcga gaaaatatga aaaacagaaa acaaagaaca gaaaaatggg tttgagagtc
     1201 agtataattt ataaaagaaa aattgtacat agaaattaac catttttgta gaagaagtta
     1261 tttttcaagc atcgttaaaa attattcaaa gcaccttatt tcatatttaa ttttaaacat
     1321 ggttaaatga acaacacggt gcgcaatcag gaaaacttga aatctgaaac tgttgttgtg
     1381 atcttcttcg caactgttca gatagcacta gtgtaatgtt aagagtgcgc gaatataatg
     1441 gaatataatg gatcacacct cctgccatca ggtaaacgtc tctgttatca catatttcca
     1501 actattaaat ttttaccttt tacagtttta catttttttg aaaaaagtaa ctttttgtct
     1561 tcaaaatccc tgacgaaaat atcaaatatt ttaatcgaga ctgcagagga accgattgat
     1621 gatttggaaa atccagcttt acctgtgtaa gaactgaaaa gtttcataac cctagggtat
     1681 tcccagttac attccccact ggctaacaat agcacccagt ttttcatcac ctttcttcaa
     1741 atttctcggc gatttgttaa aaacaaaatt tgtgtccctt ctctgatatc tctatgtctc
     1801 taaacacaag ttcatcggaa aacgaaggag ggtaggtgtt ggttgggctc ccgaagtgaa
     1861 aatagaagag caagaataga atattagaga gagagtgcag agagggcggg atagctcccg
     1921 ggattccgtt ttcttcttct ttatcttcaa cgatgatgtg tgtgcgtgtt gtatagattc
     1981 tgttgctccc ccacaactcg ctccgaaggc tcaatacaat tcaattgata ttggaggaga
     2041 gcctaccgga gtgggaggat aagaagaaac ataagaagaa gaagaagaag aagcatgctt
     2101 ctggtttttg atgctatgaa aacggcacaa aaagatgatt gaggtccctt ttcaatacct
     2161 tctctcatct ttcaaatccc attgaaacct aaaacttctc accacgcttt accattgttc
     2221 tccaaaaact tatagcaatg tctataactt ttttatctct gaaaagcagt gttccatttt
     2281 tctttttcct attttatttc aattgtttct cacatttcgt ttggattctt tgcttgtcaa
     2341 ccagcttctt cttccacttt taccgtctaa ttttcagggc agggagccat caaacccacg
     2401 accactagat ccatctagag gatccccggg attggccaaa ggacccaaag gaggaccctt
     2461 ggagggtacc ggtagaaaaa atgagtaaag gagaagaact tttcactgga gttgtcccaa
     2521 ttcttgttga attagatggt gatgttaatg ggcacaaatt ttctgtcagt ggagagggtg
     2581 aaggtgatgc aacatacgga aaacttaccc ttaaatttat ttgcactact ggaaaactac
     2641 ctgttccatg ggtaagttta aacatatata tactaactaa ccctgattat ttaaattttc
     2701 agccaacact tgtcactact ttcggttatg gtcttcaatg cttcgccaga tacccagatc
     2761 atatgaaaca gcatgacttt ttcaagagtg ccatgcccga aggttatgta caggaaagaa
     2821 ctatattttt caaagatgac gggaactaca agacacgtaa gtttaaacag ttcggtacta
     2881 actaaccata catatttaaa ttttcaggtg ctgaagtcaa gtttgaaggt gatacccttg
     2941 ttaatagaat cgagttaaaa ggtattgatt ttaaagaaga tggaaacatt cttggacaca
     3001 aattggaata caactataac tcacacaatg tatacatcat ggcagacaaa caaaagaatg
     3061 gaatcaaagt tgtaagttta aacatgattt tactaactaa ctaatctgat ttaaattttc
     3121 agaacttcaa aattagacac aacattgaag atggaagcgt tcaactagca gaccattatc
     3181 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcct
     3241 accaatctgc cctttcgaaa gatcccaacg aaaagagaga ccacatggtc cttcttgagt
     3301 ttgtaacagc tgctgggatt acacatggca tggatgaact atacaaatag cattcgtaga
     3361 attgtaagtt taaactttct catactaact aacgcgctct atttaaattt tcagaattcc
     3421 aactgagcgc cggtcgctac cattaccaac ttgtctggtg tcaaaaataa taggggccgc
     3481 tgtcatcaga gtaagtttaa actgagttct actaactaac gagtaatatt taaattttca
     3541 gcatctcgcg cccgtgcctc tgacttctaa gtccaattac tcttcaacat ccctacatgc
     3601 tctttctccc tgtgctccca ccccctattt ttgttattat caaaaaactt ctcttaattt
     3661 ctttgttttt tagcttcttt taagtcacct ctaacaatga aattgtgtag attcaaaaat
     3721 agaattaatt cgtaataaaa agtcgaaaaa aattgtgctc cctcccccca ttaataataa
     3781 ttctatccca aaatctacac aatgttctgt gtacacttct tatgtttttt acttctgata
     3841 aatttttttg aaacatcata gaaaaaaccg cacacaaaat accttatcat atgttacgtt
     3901 tcagtttatg accgcaattt ttatttcttc gcacgtctgg gcctctcatg acgtcaaatc
     3961 atgctcatcg tgaaaaagtt ttggagtatt tttggaattt ttcaatcaag tgaaagttta
     4021 tgaaattaat tttcctgctt ttgctttttg gggtttcccc tattgtttgt caagatttcg
     4081 aggacggcgt ttttcttgct aaaatcacaa gtattgatga gcacgatgca agaaagatcg
     4141 gaagaaggtt tgggtttgag gctcagtgga aggtgagtag aagttgataa tttgaaagtg
     4201 gagtagtgtc tatggggttt ttgccttaaa tgacagaata cattcccaat ataccaaaca
     4261 taactgtttc ctactagtcg gccgtacggg ccctttcgtc tcgcgcgttt cggtgatgac
     4321 ggtgaaaacc tctgacacat gcagctcccg gagacggtca cagcttgtct gtaagcggat
     4381 gccgggagca gacaagcccg tcagggcgcg tcagcgggtg ttggcgggtg tcggggctgg
     4441 cttaactatg cggcatcaga gcagattgta ctgagagtgc accatatgcg gtgtgaaata
     4501 ccgcacagat gcgtaaggag aaaataccgc atcaggcggc cttaagggcc tcgtgatacg
     4561 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     4621 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     4681 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     4741 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4801 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4861 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4921 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4981 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     5041 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     5101 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     5161 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     5221 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     5281 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     5341 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     5401 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     5461 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     5521 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     5581 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     5641 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     5701 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     5761 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5821 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5881 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5941 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     6001 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     6061 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     6121 gcgaacgacc tacaccgaac tgagatacct acagcgtgag cattgagaaa gcgccacgct
     6181 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     6241 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     6301 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     6361 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     6421 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     6481 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     6541 gcgcccaata cgcaaaccgc ctctccccgc gcgttggccg attcattaat gcagctggca
     6601 cgacaggttt cccgactgga aagcgggcag tgagcgcaac gcaattaatg tgagttagct
     6661 cactcattag gcaccccagg ctttacactt tatgcttccg gctcgtatgt tgtgtggaat
     6721 tgtgagcgga taacaatttc acacaggaaa cagct
//
