LOCUS       sequence-8371-ex        2931 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 2931)
  TITLE     Purification free plasmid
REFERENCE   2  (bases 1 to 2931)
  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: pGT3
FEATURES             Location/Qualifiers
     source          1..2931
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     protein_bind    52..73
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(88..93,94..111,112..118)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    126..142
                     /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     131..153
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     150..166
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     150..166
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             234..950
                     /codon_start=1
                     /product="GFP variant optimized for excitation by UV light"
                     /label=GFPuv
                     /translation="MSKGEELFTGVVPILVELDGDVNGHKFSVSGEGEGDATYGKLTLK
                     FICTTGKLPVPWPTLVTTLTYGVQCFSRYPDHMKRHDFFKSAMPEGYVQERTISFKDDG
                     NYKTRAEVKFEGDTLVNRIELKGIDFKEDGNILGHKLEYNYNSHNVYITADKQKNGIKA
                     NFKIRHNIEDGSVQLADHYQQNTPIGDGPVLLPDNHYLSTQSALLKDPNEKRDHMVLLE
                     FVTAAGITHGMDELYK"
     primer_bind     complement(264..292)
                     /label=GFP-R
                     /note="GFP, reverse primer. Does NOT anneal to EGFP"
     primer_bind     887..907
                     /label=GFP-F
                     /note="GFP, forward primer. Does NOT anneal to EGFP"
     promoter        1010..1114
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1115..1183,1184..1975)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(1333..1352)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2146..2734
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2635..2654
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2888..2905
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 ctggcacgac aggtttcccg actggaaagc gggcagtgag cgcaacgcaa ttaatgtgag
       61 ttagctcact cattaggcac cccaggcttt acactttatg cttccggctc gtatgttgtg
      121 tggaattgtg agcggataac aatttcacac aggaaacagc tatgaccatg attacgccaa
      181 gcttgcatgc ctgcaggtcg actctagagg atccccgggt accggtagaa aaaatgagta
      241 aaggagaaga acttttcact ggagttgtcc caattcttgt tgaattagat ggtgatgtta
      301 atgggcacaa attttctgtc agtggagagg gtgaaggtga tgcaacatac ggaaaactta
      361 cccttaaatt tatttgcact actggaaaac tacctgttcc atggccaaca cttgtcacta
      421 ctctgacgta tggtgttcaa tgcttttccc gttatccgga tcatatgaaa cggcatgact
      481 ttttcaagag tgccatgccc gaaggttatg tacaggaacg cacgatatct ttcaaagatg
      541 acgggaacta caagacgcgt gctgaagtca agtttgaagg tgataccctt gttaatcgta
      601 tcgagttaaa aggtattgat tttaaagaag atggaaacat tctcggacac aaactcgagt
      661 acaactataa ctcacacaat gtatacatca cggcagacaa acaaaagaat ggaatcaaag
      721 ctaacttcaa aattcgccac aacattgaag atggatccgt tcaactagca gaccattatc
      781 aacaaaatac tccaattggc gatggccctg tccttttacc agacaaccat tacctgtcga
      841 cacaatctgc ccttttgaaa gatcccaacg aaaagcgtga ccacatggtc cttcttgagt
      901 ttgtaactgc tgctgggatt acacatggca tggatgagct ctacaaataa tgaattccca
      961 gactagatct ggcgcgccga cgtcaggtgg cacttttcgg ggaaatgtgc gcggaacccc
     1021 tatttgttta tttttctaaa tacattcaaa tatgtatccg ctcatgagac aataaccctg
     1081 ataaatgctt caataatatt gaaaaaggaa gagtatgagt attcaacatt tccgtgtcgc
     1141 ccttattccc ttttttgcgg cattttgcct tcctgttttt gctcacccag aaacgctggt
     1201 gaaagtaaaa gatgctgaag atcagttggg tgcacgagtg ggttacatcg aactggatct
     1261 caacagcggt aagatccttg agagttttcg ccccgaagaa cgttttccaa tgatgagcac
     1321 ttttaaagtt ctgctatgtg gcgcggtatt atcccgtgtt gacgccgggc aagagcaact
     1381 cggtcgccgc atacactatt ctcagaatga cttggttgag tactcaccag tcacagaaaa
     1441 gcatcttacg gatggcatga cagtaagaga attatgcagt gctgccataa ccatgagtga
     1501 taacactgcg gccaacttac ttctgacaac gatcggagga ccgaaggagc taaccgcttt
     1561 tttgcacaac atgggggatc atgtaactcg ccttgatcgt tgggaaccgg agctgaatga
     1621 agccatacca aacgacgagc gtgacaccac gatgcctgca gcaatggcaa caacgttgcg
     1681 caaactatta actggcgaac tacttactct agcttcccgg caacaattaa tagactggat
     1741 ggaggcggat aaagttgcag gaccacttct gcgctcggcc cttccggctg gctggtttat
     1801 tgctgataaa tctggagccg gtgagcgtgg gtctcgcggt atcattgcag cactggggcc
     1861 agatggtaag ccctcccgta tcgtagttat ctacacgacg gggagtcagg caactatgga
     1921 tgaacgaaat agacagatcg ctgagatagg tgcgtcactg attaagcatt ggtaactgtc
     1981 agaccaagtt tactcatata tactttagat tgatttaaaa cttcattttt aatttaaaag
     2041 gatctaggtg aagatccttt ttgataatct catgaccaaa atcccttaac gtgagttttc
     2101 gttccactga gcgtcagacc ccgtagaaaa gatcaaagga tcttcttgag atcctttttt
     2161 tctgcgcgta atctgctgct tgcaaacaaa aaaaccaccg ctaccagcgg tggtttgttt
     2221 gccggatcaa gagctaccaa ctctttttcc gaaggtaact ggcttcagca gagcgcagat
     2281 accaaatact gtccttctag tgtagccgta gttaggccac cacttcaaga actctgtagc
     2341 accgcctaca tacctcgctc tgctaatcct gttaccagtg gctgctgcca gtggcgataa
     2401 gtcgtgtctt accgggttgg actcaagacg atagttaccg gataaggcgc agcggtcggg
     2461 ctgaacgggg ggttcgtgca cacagcccag cttggagcga acgacctaca ccgaactgag
     2521 atacctacag cgtgagctat gagaaagcgc cacgcttccc gaagggagaa aggcggacag
     2581 gtatccggta agcggcaggg tcggaacagg agagcgcacg agggagcttc cagggggaaa
     2641 cgcctggtat ctttatagtc ctgtcgggtt tcgccacctc tgacttgagc gtcgattttt
     2701 gtgatgctcg tcaggggggc ggagcctatg gaaaaacgcc agcaacgcgg cctttttacg
     2761 gttcctggcc ttttgctggc cttttgctca catgttcttt cctgcgttat cccctgattc
     2821 tgtggataac cgtattaccg cctttgagtg agctgatacc gctcgccgca gccgaacgac
     2881 cgagcgcagc gagtcagtga gcgaggaagc ggcgcgccat gcatacgttg g
//
