LOCUS       sequence-7434-ex        3267 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 3267)
  AUTHORS   Park HK, Zeng C
  TITLE     Construction of an XcmI-generated T vector bearing green fluorescent
            protein marker for direct cloning of PCR products.
  JOURNAL   Anal Biochem. 2007 Jan 1;360(1):144-5. doi: 
            10.1016/j.ab.2006.10.031. Epub 2006 Nov 7.
  PUBMED    17113027
REFERENCE   2  (bases 1 to 3267)
  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"; doi: 
            "10.1016/j.ab.2006.10.031"; journalName: "Anal Biochem"; date: 
            "2007-01-1- 1"; volume: "360"; issue: "1"; pages: "144-5"
COMMENT     Sequence Label: pGT2
FEATURES             Location/Qualifiers
     source          1..3267
                     /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..947
                     /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"
     primer_bind     complement(990..1008)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1076..1180
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1181..1249,1250..2041)
                     /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(1399..1418)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2212..2800
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2701..2720
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2954..2971
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2986..3126
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3052..3071)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     3212..3234
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse 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 atcccagggt tccggtggaa 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 ctacaaacca cactagtgct
      961 ggcggccgcg aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
     1021 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
     1081 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
     1141 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
     1201 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
     1261 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
     1321 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
     1381 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1441 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1501 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1561 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1621 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1681 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     1741 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1801 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1861 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1921 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1981 tatggatgaa cgaaatagac agatcgctga gataggtgcg tcactgatta agcattggta
     2041 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     2101 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     2161 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     2221 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     2281 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     2341 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     2401 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2461 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2521 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2581 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2641 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2701 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2761 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2821 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2881 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2941 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     3001 tctccttacg catctgtgcg gtatttcaca ccgcatatgg tgcactctca gtacaatctg
     3061 ctctgatgcc gcatagttaa gccagtatac actccgctat cgctacgtga ctgggtcatg
     3121 gctgcgcccc gacacccgcc aacacccgct gacgcgccct gacgggcttg tctgctcccg
     3181 gcatccgctt acagacaagc tgtgaccgtc tccgggagct gcatgtgtca gaggttttca
     3241 ccgtcatcac cgaaacgcgc gaggcag
//
