LOCUS       Exported                5290 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 5290)
  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..5290
                     /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..857
                     /codon_start=1
                     /label=RAP1A_G12C_L96F_CL
                     /translation="MREYKLVVLGSCGVGKSALTVQFVQGIFVEKYDPTIEDSYRKQVE
                     VDSQQSMLEILDTAGTEQFTAMRDLYMKNGQGFALVYSITAQSTFNDLQDFREQILRVK
                     DTEDVPMILVGNKSDLEDERVVGKEQGQNLARQWSNSAFLESSAKSKINVNEIFYDLVR
                     QINRK"
     terminator      1149..1235
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      1327..1354
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
     promoter        1374..1465
                     /gene="bla"
                     /label=AmpR promoter
     CDS             1466..2326
                     /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      2497..3085
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     rep_origin      complement(3333..3761)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        4091..4168
                     /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             4169..5251
                     /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 atgagggaat
      361 ataaattagt tgtactagga tcctgcggtg taggcaagtc cgcgttgacc gttcagttcg
      421 tgcagggcat tttcgtggag aaatacgacc cgaccattga agacagctat cgtaagcagg
      481 ttgaggtgga ttctcagcag agcatgctcg agatcctgga tacggcaggc accgagcaat
      541 ttacggctat gcgtgattta tatatgaaaa acggtcaagg ctttgccctg gtctacagca
      601 ttaccgcgca gtcgaccttc aacgatctgc aggatttcag agaacagatc ctgcgcgtga
      661 aggacactga ggacgttccg atgattctgg tcggtaataa gagtgatttg gaggacgaac
      721 gtgttgttgg taaagaacaa ggtcaaaacc tggcgcgtca atggagcaat agcgcgtttc
      781 tggaatctag cgctaaaagc aaaatcaacg tgaatgaaat cttctacgac ctggtgcgtc
      841 aaatcaaccg caagtaagaa ttcaaaggcc tacgtcgacg agctcactag tcgcggccgc
      901 tttcgaatct agagcctgca gtctcgaggc atgcggtacc aagcttggct gttttggcgg
      961 atgagagaag attttcagcc tgatacagat taaatcagaa cgcagaagcg gtctgataaa
     1021 acagaatttg cctggcggca gtagcgcggt ggtcccacct gaccccatgc cgaactcaga
     1081 agtgaaacgc cgtagcgccg atggtagtgt ggggtctccc catgcgagag tagggaactg
     1141 ccaggcatca aataaaacga aaggctcagt cgaaagactg ggcctttcgt tttatctgtt
     1201 gtttgtcggt gaacgctctc ctgagtagga caaatccgcc gggagcggat ttgaacgttg
     1261 cgaagcaacg gcccggaggg tggcgggcag gacgcccgcc ataaactgcc aggcatcaaa
     1321 ttaagcagaa ggccatcctg acggatggcc tttttgcgtt tctacaaact ctttttgttt
     1381 atttttctaa atacattcaa atatgtatcc gctcatgaga caataaccct gataaatgct
     1441 tcaataatat tgaaaaagga agagtatgag tattcaacat ttccgtgtcg cccttattcc
     1501 cttttttgcg gcattttgcc ttcctgtttt tgctcaccca gaaacgctgg tgaaagtaaa
     1561 agatgctgaa gatcagttgg gtgcacgagt gggttacatc gaactggatc tcaacagcgg
     1621 taagatcctt gagagttttc gccccgaaga acgttttcca atgatgagca cttttaaagt
     1681 tctgctatgt ggcgcggtat tatcccgtgt tgacgccggg caagagcaac tcggtcgccg
     1741 catacactat tctcagaatg acttggttga gtactcacca gtcacagaaa agcatcttac
     1801 ggatggcatg acagtaagag aattatgcag tgctgccata accatgagtg ataacactgc
     1861 ggccaactta cttctgacaa cgatcggagg accgaaggag ctaaccgctt ttttgcacaa
     1921 catgggggat catgtaactc gccttgatcg ttgggaaccg gagctgaatg aagccatacc
     1981 aaacgacgag cgtgacacca cgatgcctac agcaatggca acaacgttgc gcaaactatt
     2041 aactggcgaa ctacttactc tagcttcccg gcaacaatta atagactgga tggaggcgga
     2101 taaagttgca ggaccacttc tgcgctcggc ccttccggct ggctggttta ttgctgataa
     2161 atctggagcc ggtgagcgtg ggtctcgcgg tatcattgca gcactggggc cagatggtaa
     2221 gccctcccgt atcgtagtta tctacacgac ggggagtcag gcaactatgg atgaacgaaa
     2281 tagacagatc gctgagatag gtgcctcact gattaagcat tggtaactgt cagaccaagt
     2341 ttactcatat atactttaga ttgatttaaa acttcatttt taatttaaaa ggatctaggt
     2401 gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt cgttccactg
     2461 agcgtcagac cccgtagaaa agatcaaagg atcttcttga gatccttttt ttctgcgcgt
     2521 aatctgctgc ttgcaaacaa aaaaaccacc gctaccagcg gtggtttgtt tgccggatca
     2581 agagctacca actctttttc cgaaggtaac tggcttcagc agagcgcaga taccaaatac
     2641 tgtccttcta gtgtagccgt agttaggcca ccacttcaag aactctgtag caccgcctac
     2701 atacctcgct ctgctaatcc tgttaccagt ggctgctgcc agtggcgata agtcgtgtct
     2761 taccgggttg gactcaagac gatagttacc ggataaggcg cagcggtcgg gctgaacggg
     2821 gggttcgtgc acacagccca gcttggagcg aacgacctac accgaactga gatacctaca
     2881 gcgtgagcta tgagaaagcg ccacgcttcc cgaagggaga aaggcggaca ggtatccggt
     2941 aagcggcagg gtcggaacag gagagcgcac gagggagctt ccagggggaa acgcctggta
     3001 tctttatagt cctgtcgggt ttcgccacct ctgacttgag cgtcgatttt tgtgatgctc
     3061 gtcagggggg cggagcctat ggaaaaacgc cagcaacgcg gcctttttac ggttcctggc
     3121 cttttgctgg ccttttgctc acatgttctt tcctgcgtta tcccctgatt ctgtggataa
     3181 ccgtattacc gcctttgagt gagctgatac cgctcgccgc agccgaacga ccgagcgcag
     3241 cgagtcagtg agcgaggaag cggaagagcg cctgatgcgg tattttctcc ttacgcatct
     3301 gtgcggtatt tcacaccgca taattttgtt aaaattcgcg ttaaattttt gttaaatcag
     3361 ctcatttttt aaccaatagg ccgaaatcgg caaaatccct tataaatcaa aagaatagac
     3421 cgagataggg ttgagtgttg ttccagtttg gaacaagagt ccactattaa agaacgtgga
     3481 ctccaacgtc aaagggcgaa aaaccgtcta tcagggcgat ggcccactac gtgaaccatc
     3541 accctaatca agttttttgg ggtcgaggtg ccgtaaagca ctaaatcgga accctaaagg
     3601 gagcccccga tttagagctt gacggggaaa gccggcgaac gtggcgagaa aggaagggaa
     3661 gaaagcgaaa ggagcgggcg ctagggcgct ggcaagtgta gcggtcacgc tgcgcgtaac
     3721 caccacaccc gccgcgctta atgcgccgct acagggcgcg tcccattcgc cattcaggct
     3781 gctatggtgc actctcagta caatctgctc tgatgccgca tagttaagcc agtaccagtc
     3841 acgtagcgat atcgctacgt gactggtaca ctccgctatc gctacgtgac tgggtcatgg
     3901 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     3961 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     4021 cgtcatcacc gaaacgcgcg aggcagcaga tcaattcgcg cgcgaaggcg aagcggcatg
     4081 catttacgtt gacaccatcg aatggtgcaa aacctttcgc ggtatggcat gatagcgccc
     4141 ggaagagagt caattcaggg tggtgaatgt gaaaccagta acgttatacg atgtcgcaga
     4201 gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg aaccaggcca gccacgtttc
     4261 tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag ctgaattaca ttcccaaccg
     4321 cgtggcacaa caactggcgg gcaaacagtc gttgctgatt ggcgttgcca cctccagtct
     4381 ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa tctcgcgccg atcaactggg
     4441 tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc gaagcctgta aagcggcggt
     4501 gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt aactatccgc tggatgacca
     4561 ggatgccatt gctgtggaag ctgcctgcac taatgttccg gcgttatttc ttgatgtctc
     4621 tgaccagaca cccatcaaca gtattatttt ctcccatgaa gacggtacgc gactgggcgt
     4681 ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg ttagcgggcc cattaagttc
     4741 tgtctcggcg cgtctgcgtc tggctggctg gcataaatat ctcactcgca atcaaattca
     4801 gccgatagcg gaacgggaag gcgactggag tgccatgtcc ggttttcaac aaaccatgca
     4861 aatgctgaat gagggcatcg ttcccactgc gatgctggtt gccaacgatc agatggcgct
     4921 gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt ggtgcggata tctcggtagt
     4981 gggatacgac gataccgaag acagctcatg ttatatcccg ccgttaacca ccatcaaaca
     5041 ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg ctgcaactct ctcagggcca
     5101 ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg aaaagaaaaa ccaccctggc
     5161 acccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc agctggcacg
     5221 acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgta agttagcgcg
     5281 aattgatctg
//
