LOCUS       sequence-442880-        5298 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 5298)
  AUTHORS   Chapman S, Faulkner C, Kaiserli E, Garcia-Mata C, Savenkov EI, 
            Roberts AG, Oparka KJ, Christie JM
  TITLE     The photoreversible fluorescent protein iLOV outperforms GFP as a 
            reporter of plant virus infection.
  JOURNAL   Proc Natl Acad Sci U S A. 2008 Dec 16. 105(50):20038-43.
  PUBMED    19060199
REFERENCE   2  (bases 1 to 5298)
  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: "Proc Natl 
            Acad Sci U S A. 2008 Dec 16. 105(50):20038-43."
COMMENT     Sequence Label: pGEX iLOV
FEATURES             Location/Qualifiers
     source          1..5298
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        59..136
                     /gene="lacI"
                     /label=lacI promoter
     CDS             137..1219
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /label=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)."
                     /translation="MKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     primer_bind     complement(156..175)
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        join(1268..1273,1274..1291,1292..1298)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1306..1322
                     /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     1330..1346
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     1330..1346
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     CDS             1342..1869
                     /codon_start=1
                     /gene="lacZ fragment"
                     /product="LacZ-alpha fragment of beta-galactosidase"
                     /label=lacZ-alpha
                     /translation="MTMITDSLAVVLQRRDWENPGVTQLNRLAAHPPFASWRNSEEART
                     DRPSQQLRSLNGEWRFAWFPAPEAVPESWLECDLPEADTVVVPSNWQMHGYDAPIYTNV
                     TYPITVNPPFVPTENPTGCYSLTFNVDESWLQEGQTRIIFDGVGISLSTARCTNASGVR
                     QPSEAVVWLCRS"
     primer_bind     complement(1362..1379)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1362..1378)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1371..1393)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     promoter        join(1971..1976,1977..1992,1993..1999)
                     /label=tac promoter
                     /note="strong E. coli promoter; hybrid between the trp and 
                     lac UV5 promoters"
     protein_bind    2007..2023
                     /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)."
     CDS             2046..2699
                     /codon_start=1
                     /product="glutathione S-transferase from Schistosoma 
                     japonicum"
                     /label=GST
                     /translation="MSPILGYWKIKGLVQPTRLLLEYLEEKYEEHLYERDEGDKWRNKK
                     FELGLEFPNLPYYIDGDVKLTQSMAIIRYIADKHNMLGGCPKERAEISMLEGAVLDIRY
                     GVSRIAYSKDFETLKVDFLSKLPEMLKMFEDRLCHKTYLNGDHVTHPDFMLYDALDVVL
                     YMDPMCLDAFPKLVCFKKRIEAIPQIDKYLKSSKYIAWPLQGWQATFGGGDHPPK"
     primer_bind     2657..2679
                     /label=pGEX 5'
                     /note="pGEX vectors, Glutathione-S-transferase, forward 
                     primer"
     CDS             2706..2723
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(3136..3158)
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(3303..3321)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        3389..3493
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3494..3562,3563..4354)
                     /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(3712..3731)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4525..5113
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     5014..5033
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5267..5284
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 cctgatgcgg tattttctcc ttacgcatct gtgcggtatt tcacaccgca taaattccga
       61 caccatcgaa tggcgcaaaa cctttcgcgg tatggcatga tagcgcccgg aagagagtca
      121 attcagggtg gtgaatgtga aaccagtaac gttatacgat gtcgcagagt atgccggtgt
      181 ctcttatcag accgtttccc gcgtggtgaa ccaggccagc cacgtttctg cgaaaacgcg
      241 ggaaaaagtg gaagcggcga tggcggagct gaattacatt cccaaccgcg tggcacaaca
      301 actggcgggc aaacagtcgt tgctgattgg cgttgccacc tccagtctgg ccctgcacgc
      361 gccgtcgcaa attgtcgcgg cgattaaatc tcgcgccgat caactgggtg ccagcgtggt
      421 ggtgtcgatg gtagaacgaa gcggcgtcga agcctgtaaa gcggcggtgc acaatcttct
      481 cgcgcaacgc gtcagtgggc tgatcattaa ctatccgctg gatgaccagg atgccattgc
      541 tgtggaagct gcctgcacta atgttccggc gttatttctt gatgtctctg accagacacc
      601 catcaacagt attattttct cccatgaaga cggtacgcga ctgggcgtgg agcatctggt
      661 cgcattgggt caccagcaaa tcgcgctgtt agcgggccca ttaagttctg tctcggcgcg
      721 tctgcgtctg gctggctggc ataaatatct cactcgcaat caaattcagc cgatagcgga
      781 acgggaaggc gactggagtg ccatgtccgg ttttcaacaa accatgcaaa tgctgaatga
      841 gggcatcgtt cccactgcga tgctggttgc caacgatcag atggcgctgg gcgcaatgcg
      901 cgccattacc gagtccgggc tgcgcgttgg tgcggatatc tcggtagtgg gatacgacga
      961 taccgaagac agctcatgtt atatcccgcc gttaaccacc atcaaacagg attttcgcct
     1021 gctggggcaa accagcgtgg accgcttgct gcaactctct cagggccagg cggtgaaggg
     1081 caatcagctg ttgcccgtct cactggtgaa aagaaaaacc accctggcgc ccaatacgca
     1141 aaccgcctct ccccgcgcgt tggccgattc attaatgcag ctggcacgac aggtttcccg
     1201 actggaaagc gggcagtgag cgcaacgcaa ttaatgtaag ttagctcact cattaggcac
     1261 cccaggcttt acactttatg cttccggctc gtatgttgtg tggaattgtg agcggataac
     1321 aatttcacac aggaaacagc tatgaccatg attacggatt cactggccgt cgttttacaa
     1381 cgtcgtgact gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct
     1441 ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc
     1501 agcctgaatg gcgaatggcg ctttgcctgg tttccggcac cagaagcggt gccggaaagc
     1561 tggctggagt gcgatcttcc tgaggccgat actgtcgtcg tcccctcaaa ctggcagatg
     1621 cacggttacg atgcgcccat ctacaccaac gtgacctatc ccattacggt caatccgccg
     1681 tttgttccca cggagaatcc gacgggttgt tactcgctca catttaatgt tgatgaaagc
     1741 tggctacagg aaggccagac gcgaattatt tttgatggcg ttggaattag cttatcgact
     1801 gcacggtgca ccaatgcttc tggcgtcagg cagccatcgg aagctgtggt atggctgtgc
     1861 aggtcgtaaa tcactgcata attcgtgtcg ctcaaggcgc actcccgttc tggataatgt
     1921 tttttgcgcc gacatcataa cggttctggc aaatattctg aaatgagctg ttgacaatta
     1981 atcatcggct cgtataatgt gtggaattgt gagcggataa caatttcaca caggaaacag
     2041 tattcatgtc ccctatacta ggttattgga aaattaaggg ccttgtgcaa cccactcgac
     2101 ttcttttgga atatcttgaa gaaaaatatg aagagcattt gtatgagcgc gatgaaggtg
     2161 ataaatggcg aaacaaaaag tttgaattgg gtttggagtt tcccaatctt ccttattata
     2221 ttgatggtga tgttaaatta acacagtcta tggccatcat acgttatata gctgacaagc
     2281 acaacatgtt gggtggttgt ccaaaagagc gtgcagagat ttcaatgctt gaaggagcgg
     2341 ttttggatat tagatacggt gtttcgagaa ttgcatatag taaagacttt gaaactctca
     2401 aagttgattt tcttagcaag ctacctgaaa tgctgaaaat gttcgaagat cgtttatgtc
     2461 ataaaacata tttaaatggt gatcatgtaa cccatcctga cttcatgttg tatgacgctc
     2521 ttgatgttgt tttatacatg gacccaatgt gcctggatgc gttcccaaaa ttagtttgtt
     2581 ttaaaaaacg tattgaagct atcccacaaa ttgataagta cttgaaatcc agcaagtata
     2641 tagcatggcc tttgcagggc tggcaagcca cgtttggtgg tggcgaccat cctccaaaat
     2701 cggatctggt tccgcgtgga tccccggaat tcatagagaa gaatttcgtc atcactgatc
     2761 ctaggcttcc cgataatccc attatctttg catcagacgg ctttcttgaa ttgacagagt
     2821 attcgcgcga ggaaatattg gggagaaatg cccggtttct tcaggggcca gagacagatc
     2881 aagcgactgt ccagaagata agagacgcaa ttagagatca gagggagact actgtgcagt
     2941 tgataaacta cactaaaagc ggaaagaaat tctggaactt actccacctg caacctgtgc
     3001 gtgatcagaa gggagagctt caatacttca tcggtgtgca gctcgatgga agtgatcatg
     3061 tataagtcga ctcgagcggc cgcatcgtga ctgactgacg atctgcctcg cgcgtttcgg
     3121 tgatgacggt gaaaacctct gacacatgca gctcccggag acggtcacag cttgtctgta
     3181 agcggatgcc gggagcagac aagcccgtca gggcgcgtca gcgggtgttg gcgggtgtcg
     3241 gggcgcagcc atgacccagt cacgtagcga tagcggagtg tataattctt gaagacgaaa
     3301 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     3361 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     3421 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     3481 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     3541 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     3601 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     3661 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     3721 cgcggtatta tcccgtgttg acgccgggca agagcaactc ggtcgccgca tacactattc
     3781 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     3841 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     3901 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     3961 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     4021 tgacaccacg atgcctgcag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     4081 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     4141 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     4201 tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     4261 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     4321 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     4381 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     4441 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     4501 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     4561 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     4621 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     4681 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     4741 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     4801 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     4861 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     4921 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     4981 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     5041 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     5101 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     5161 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     5221 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     5281 cgaggaagcg gaagagcg
//
