LOCUS       sequence-361543-        3139 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Greengate entry vector pGGC000 modified to contain bladderwort 
            insulator 1.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 3139)
  AUTHORS   Park JY, Kovar L, LaFayette PR, Wallace JG, Parrott WA
  TITLE     Plant-derived insulator-like sequences for control of transgene 
            expression
  JOURNAL   bioRxiv 2021.11.04.467280
REFERENCE   2  (bases 1 to 3139)
  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"; journalName: "bioRxiv 
            2021.11.04.467280"
COMMENT     Sequence Label: CD Ugi1
FEATURES             Location/Qualifiers
     source          1..3139
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(3..22)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(4..22)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(32..54)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(248..267)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     367..389
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(427..445)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        513..617
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(618..686,687..1478)
                     /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(836..855)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1649..2237
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2138..2157
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2391..2408
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2525..2546
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(2561..2566,2567..2584,2585..2591)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2599..2615
                     /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)."
     promoter        2629..2647
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     2629..2646
                     /label=SP6
                     /note="SP6 promoter, forward primer"
ORIGIN
        1 cgccctatag tgagtcgtat tacctggccg tcgttttaca acgtcgtgac tgggaaaacc
       61 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
      121 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
      181 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atatggtgca
      241 ctctcagtac aatctgctct gatgccgcat agttaagcca gccccgacac ccgccaacac
      301 ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc cgcttacaga caagctgtga
      361 ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc atcaccgaaa cgcgcgagac
      421 gaaagggcct cgtgatacgc ctatttttat aggttaatgt catgataata atggtttctt
      481 agacgtcagg tggcactttt cggggaaatg tgcgcggaac ccctatttgt ttatttttct
      541 aaatacattc aaatatgtat ccgctcatga gacaataacc ctgataaatg cttcaataat
      601 attgaaaaag gaagagtatg agtattcaac atttccgtgt cgcccttatt cccttttttg
      661 cggcattttg ccttcctgtt tttgctcacc cagaaacgct ggtgaaagta aaagatgctg
      721 aagatcagtt gggtgcacga gtgggttaca tcgaactgga tctcaacagc ggtaagatcc
      781 ttgagagttt tcgccccgaa gaacgttttc caatgatgag cacttttaaa gttctgctat
      841 gtggcgcggt attatcccgt attgacgccg ggcaagagca actcggtcgc cgcatacact
      901 attctcagaa tgacttggtt gagtactcac cagtcacaga aaagcatctt acggatggca
      961 tgacagtaag agaattatgc agtgctgcca taaccatgag tgataacact gcggccaact
     1021 tacttctgac aacgatcgga ggaccgaagg agctaaccgc ttttttgcac aacatggggg
     1081 atcatgtaac tcgccttgat cgttgggaac cggagctgaa tgaagccata ccaaacgacg
     1141 agcgtgacac cacgatgcct gtagcaatgg caacaacgtt gcgcaaacta ttaactggcg
     1201 aactacttac tctagcttcc cggcaacaat taatagactg gatggaggcg gataaagttg
     1261 caggaccact tctgcgctcg gcccttccgg ctggctggtt tattgctgat aaatctggag
     1321 ccggtgagcg tggctctcga ggtatcattg cagcactggg gccagatggt aagccctccc
     1381 gtatcgtagt tatctacacg acggggagtc aggcaactat ggatgaacga aatagacaga
     1441 tcgctgagat aggtgcctca ctgattaagc attggtaact gtcagaccaa gtttactcat
     1501 atatacttta gattgattta aaacttcatt tttaatttaa aaggatctag gtgaagatcc
     1561 tttttgataa tctcatgacc aaaatccctt aacgtgagtt ttcgttccac tgagcgtcag
     1621 accccgtaga aaagatcaaa ggatcttctt gagatccttt ttttctgcgc gtaatctgct
     1681 gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg tttgccggat caagagctac
     1741 caactctttt tccgaaggta actggcttca gcagagcgca gataccaaat actgttcttc
     1801 tagtgtagcc gtagttaggc caccacttca agaactctgt agcaccgcct acatacctcg
     1861 ctctgctaat cctgttacca gtggctgctg ccagtggcga taagtcgtgt cttaccgggt
     1921 tggactcaag acgatagtta ccggataagg cgcagcggtc gggctgaacg gggggttcgt
     1981 gcacacagcc cagcttggag cgaacgacct acaccgaact gagataccta cagcgtgagc
     2041 tatgagaaag cgccacgctt cccgaaggga gaaaggcgga caggtatccg gtaagcggca
     2101 gggtcggaac aggagagcgc acgagggagc ttccaggggg aaacgcctgg tatctttata
     2161 gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt tttgtgatgc tcgtcagggg
     2221 ggcggagcct atggaaaaac gccagcaacg cggccttttt acggttcctg gccttttgct
     2281 ggccttttgc tcacatgttc tttcctgcgt tatcccctga ttctgtggat aaccgtatta
     2341 ccgcctttga gtgagctgat accgctcgcc gcagccgaac gaccgagcgc agcgagtcag
     2401 tgagcgagga agcggaagag cgcccaatac gcaaaccgcc tctccccgcg cgttggccga
     2461 ttcattaatg cagctggcac gacaggtttc ccgactggaa agcgggcagt gagcgcaacg
     2521 caattaatgt gagttagctc actcattagg caccccaggc tttacacttt atgcttccgg
     2581 ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca cacgagctat ttaggtgaca
     2641 ctatagaagt gaagcttggt ctcaggctcc cttaatttct tctttctttt ttttttccct
     2701 tttcctgtaa ataaatgttg caccaattac aacttacaac cccaagctta aagaaagaga
     2761 tcttctccaa tttgctggct cagcagccgc aaaacctacc tcataacacg aggcttgaag
     2821 gggcagggca gcacacaaca cttgggaggt gcatcgtcta ttactcaaca ggacaactta
     2881 ttctagcagc aatcatgttg agatttcagg tggataaatc caaagaaaga ttggcgacca
     2941 tcatcccact gagggagaga acacccaccc agagtgatga ttatccaagg catcatgttg
     3001 tggtttcgac cgtattgcaa tagcaagtat ctcttccgta cgaatcatca tcccccgtgg
     3061 gaggcagtga gttgttattc gtggtgatgc agctgagttt gtgtcgtcca ctgatcctct
     3121 ttcagtgaga ccgaattct
//
