LOCUS       Exported                5317 bp ds-DNA     circular SYN 15-JUL-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5317)
  AUTHORS   Trial User
  TITLE     Direct Submission
  JOURNAL   Exported Wednesday, Jul 17, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5317
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    230..246
                     /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             279..296
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             318..338
                     /codon_start=1
                     /product="tobacco etch virus (TEV) protease recognition and
                     cleavage site"
                     /label=TEV site
                     /translation="ENLYFQG"
     CDS             351..884
                     /codon_start=1
                     /label=Rac1
                     /translation="MQAIKSVVVGDCAVGKTSLLISYTTNAFPGEYIPTVFDNYSANVM
                     VDGKPVNLGLWDTAGQEDYDRLRPLSYPQTDVFLISFSLVSPASFENVRAKWYPEVRHH
                     SPNTPIILVGTKLDLRDDKDTIEKLKEKKLTPITYPQGLAMAKEIGAVKYLESSALTQR
                     GLKTVFDEAIRAVL"
     terminator      1176..1262
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1354..1381
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1401..1492
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1493..2353
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPTAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      2524..3112
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3360..3788)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4118..4195
                     /gene="lacI (mutant)"
                     /label=lacIq promoter
                     /note="In the lacIq allele, a single base change in the 
                     promoter boosts expression of the lacI gene about 10-fold."
     CDS             4196..5278
                     /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"
ORIGIN
        1 gtttgacagc ttatcatcga ctgcacggtg caccaatgct tctggcgtca ggcagccatc
       61 ggaagctgtg gtatggctgt gcaggtcgta aatcactgca taattcgtgt cgctcaaggc
      121 gcactcccgt tctggataat gttttttgcg ccgacatcat aacggttctg gcaaatattc
      181 tgaaatgagc tgttgacaat taatcatccg gtccgtataa tctgtggaat tgtgagcgga
      241 taacaatttc acacaggaaa cagaccatgt cgtactacca tcaccatcac catcacgatt
      301 acgatatccc aacgaccgaa aacctgtatt ttcagggcgc catgggatcc atgcaggcca
      361 tcaagagtgt ggtggtggga gactgtgctg taggtaaaac tagcctactg atcagttaca
      421 caaccaatgc atttcctgga gaatatatcc ctactgtctt tgacaattat tctgccaatg
      481 ttatggtaga tggaaaaccg gtgaatctgg gcttatggga tacagctgga caagaagatt
      541 atgacagatt acgcccccta tcctatccgc aaacagatgt gttcttaatt agcttttccc
      601 ttgtgagtcc tgcatcattt gaaaatgtcc gtgcaaagtg gtatcctgag gtgcggcacc
      661 acagtcccaa cactcccatc atcctagtgg gaactaaact tgatcttagg gatgataaag
      721 acacgatcga gaaactgaag gagaagaagc tgactcccat cacctatccg cagggtctag
      781 ccatggctaa ggagattggt gctgtaaaat acctggagag ctcggcgctc acacagcgag
      841 gcctcaagac agtgtttgac gaagcgatcc gagcagtcct ctgagaattc aaaggcctac
      901 gtcgacgagc tcactagtcg cggccgcttt cgaatctaga gcctgcagtc tcgaggcatg
      961 cggtaccaag cttggctgtt ttggcggatg agagaagatt ttcagcctga tacagattaa
     1021 atcagaacgc agaagcggtc tgataaaaca gaatttgcct ggcggcagta gcgcggtggt
     1081 cccacctgac cccatgccga actcagaagt gaaacgccgt agcgccgatg gtagtgtggg
     1141 gtctccccat gcgagagtag ggaactgcca ggcatcaaat aaaacgaaag gctcagtcga
     1201 aagactgggc ctttcgtttt atctgttgtt tgtcggtgaa cgctctcctg agtaggacaa
     1261 atccgccggg agcggatttg aacgttgcga agcaacggcc cggagggtgg cgggcaggac
     1321 gcccgccata aactgccagg catcaaatta agcagaaggc catcctgacg gatggccttt
     1381 ttgcgtttct acaaactctt tttgtttatt tttctaaata cattcaaata tgtatccgct
     1441 catgagacaa taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat
     1501 tcaacatttc cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc
     1561 tcacccagaa acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg
     1621 ttacatcgaa ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg
     1681 ttttccaatg atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtgttga
     1741 cgccgggcaa gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta
     1801 ctcaccagtc acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc
     1861 tgccataacc atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc
     1921 gaaggagcta accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg
     1981 ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctacagc
     2041 aatggcaaca acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca
     2101 acaattaata gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct
     2161 tccggctggc tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat
     2221 cattgcagca ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg
     2281 gagtcaggca actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat
     2341 taagcattgg taactgtcag accaagttta ctcatatata ctttagattg atttaaaact
     2401 tcatttttaa tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat
     2461 cccttaacgt gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc
     2521 ttcttgagat cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct
     2581 accagcggtg gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg
     2641 cttcagcaga gcgcagatac caaatactgt ccttctagtg tagccgtagt taggccacca
     2701 cttcaagaac tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc
     2761 tgctgccagt ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga
     2821 taaggcgcag cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac
     2881 gacctacacc gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga
     2941 agggagaaag gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag
     3001 ggagcttcca gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg
     3061 acttgagcgt cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag
     3121 caacgcggcc tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc
     3181 tgcgttatcc cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc
     3241 tcgccgcagc cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgcct
     3301 gatgcggtat tttctcctta cgcatctgtg cggtatttca caccgcataa ttttgttaaa
     3361 attcgcgtta aatttttgtt aaatcagctc attttttaac caataggccg aaatcggcaa
     3421 aatcccttat aaatcaaaag aatagaccga gatagggttg agtgttgttc cagtttggaa
     3481 caagagtcca ctattaaaga acgtggactc caacgtcaaa gggcgaaaaa ccgtctatca
     3541 gggcgatggc ccactacgtg aaccatcacc ctaatcaagt tttttggggt cgaggtgccg
     3601 taaagcacta aatcggaacc ctaaagggag cccccgattt agagcttgac ggggaaagcc
     3661 ggcgaacgtg gcgagaaagg aagggaagaa agcgaaagga gcgggcgcta gggcgctggc
     3721 aagtgtagcg gtcacgctgc gcgtaaccac cacacccgcc gcgcttaatg cgccgctaca
     3781 gggcgcgtcc cattcgccat tcaggctgct atggtgcact ctcagtacaa tctgctctga
     3841 tgccgcatag ttaagccagt accagtcacg tagcgatatc gctacgtgac tggtacactc
     3901 cgctatcgct acgtgactgg gtcatggctg cgccccgaca cccgccaaca cccgctgacg
     3961 cgccctgacg ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg
     4021 ggagctgcat gtgtcagagg ttttcaccgt catcaccgaa acgcgcgagg cagcagatca
     4081 attcgcgcgc gaaggcgaag cggcatgcat ttacgttgac accatcgaat ggtgcaaaac
     4141 ctttcgcggt atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa
     4201 accagtaacg ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg
     4261 cgtggtgaac caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat
     4321 ggcggagctg aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt
     4381 gctgattggc gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc
     4441 gattaaatct cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag
     4501 cggcgtcgaa gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct
     4561 gatcattaac tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa
     4621 tgttccggcg ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc
     4681 ccatgaagac ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat
     4741 cgcgctgtta gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca
     4801 taaatatctc actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc
     4861 catgtccggt tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat
     4921 gctggttgcc aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct
     4981 gcgcgttggt gcggatatct cggtagtggg atacgacgat accgaagaca gctcatgtta
     5041 tatcccgccg ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga
     5101 ccgcttgctg caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc
     5161 actggtgaaa agaaaaacca ccctggcacc caatacgcaa accgcctctc cccgcgcgtt
     5221 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5281 gcaacgcaat taatgtaagt tagcgcgaat tgatctg
//
