LOCUS       Exported                5781 bp ds-DNA     circular SYN 19-OCT-2024
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    Untitled
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5781)
  AUTHORS   Triple Threat
  TITLE     Direct Submission
  JOURNAL   Exported Saturday, Oct 19, 2024 from SnapGene 3.2.1
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5781
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     CDS             716..907
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /note="rop"
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     misc_feature    1009..1151
                     /note="bom"
                     /note="basis of mobility region from pBR322"
     rep_origin      complement(1337..1925)
                     /direction=LEFT
                     /note="ori"
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             2047..2862
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /note="KanR"
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     rep_origin      complement(2955..3410)
                     /direction=LEFT
                     /note="f1 ori"
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     terminator      3447..3494
                     /note="T7 terminator"
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     misc_feature    3558..4127
                     /note="YTH domain of YTHDF1"
     CDS             complement(4107..4124)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /note="6xHis"
                     /translation="HHHHHH"
     RBS             4135..4157
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     protein_bind    4172..4196
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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        complement(4197..4215)
                     /note="T7 promoter"
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        4528..4605
                     /gene="lacI"
                     /note="lacI promoter"
     CDS             4606..5688
                     /codon_start=1
                     /gene="lacI"
                     /product="lac repressor"
                     /note="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 gtgggcgcgg ggcatgacta tcgtcgccgc acttatgact gtcttcttta tcatgcaact
       61 cgtaggacag gtgccggcag cgctctgggt cattttcggc gaggaccgct ttcgctggag
      121 cgcgacgatg atcggcctgt cgcttgcggt attcggaatc ttgcacgccc tcgctcaagc
      181 cttcgtcact ggtcccgcca ccaaacgttt cggcgagaag caggccatta tcgccggcat
      241 ggcggcccca cgggtgcgca tgatcgtgct cctgtcgttg aggacccggc taggctggcg
      301 gggttgcctt actggttagc agaatgaatc accgatacgc gagcgaacgt gaagcgactg
      361 ctgctgcaaa acgtctgcga cctgagcaac aacatgaatg gtcttcggtt tccgtgtttc
      421 gtaaagtctg gaaacgcgga agtcagcgcc ctgcaccatt atgttccgga tctgcatcgc
      481 aggatgctgc tggctaccct gtggaacacc tacatctgta ttaacgaagc gctggcattg
      541 accctgagtg atttttctct ggtcccgccg catccatacc gccagttgtt taccctcaca
      601 acgttccagt aaccgggcat gttcatcatc agtaacccgt atcgtgagca tcctctctcg
      661 tttcatcggt atcattaccc ccatgaacag aaatccccct tacacggagg catcagtgac
      721 caaacaggaa aaaaccgccc ttaacatggc ccgctttatc agaagccaga cattaacgct
      781 tctggagaaa ctcaacgagc tggacgcgga tgaacaggca gacatctgtg aatcgcttca
      841 cgaccacgct gatgagcttt accgcagctg cctcgcgcgt ttcggtgatg acggtgaaaa
      901 cctctgacac atgcagctcc cggagacggt cacagcttgt ctgtaagcgg atgccgggag
      961 cagacaagcc cgtcagggcg cgtcagcggg tgttggcggg tgtcggggcg cagccatgac
     1021 ccagtcacgt agcgatagcg gagtgtatac tggcttaact atgcggcatc agagcagatt
     1081 gtactgagag tgcaccatat atgcggtgtg aaataccgca cagatgcgta aggagaaaat
     1141 accgcatcag gcgctcttcc gcttcctcgc tcactgactc gctgcgctcg gtcgttcggc
     1201 tgcggcgagc ggtatcagct cactcaaagg cggtaatacg gttatccaca gaatcagggg
     1261 ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa ggccaggaac cgtaaaaagg
     1321 ccgcgttgct ggcgtttttc cataggctcc gcccccctga cgagcatcac aaaaatcgac
     1381 gctcaagtca gaggtggcga aacccgacag gactataaag ataccaggcg tttccccctg
     1441 gaagctccct cgtgcgctct cctgttccga ccctgccgct taccggatac ctgtccgcct
     1501 ttctcccttc gggaagcgtg gcgctttctc atagctcacg ctgtaggtat ctcagttcgg
     1561 tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc ccccgttcag cccgaccgct
     1621 gcgccttatc cggtaactat cgtcttgagt ccaacccggt aagacacgac ttatcgccac
     1681 tggcagcagc cactggtaac aggattagca gagcgaggta tgtaggcggt gctacagagt
     1741 tcttgaagtg gtggcctaac tacggctaca ctagaaggac agtatttggt atctgcgctc
     1801 tgctgaagcc agttaccttc ggaaaaagag ttggtagctc ttgatccggc aaacaaacca
     1861 ccgctggtag cggtggtttt tttgtttgca agcagcagat tacgcgcaga aaaaaaggat
     1921 ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc tcagtggaac gaaaactcac
     1981 gttaagggat tttggtcatg aacaataaaa ctgtctgctt acataaacag taatacaagg
     2041 ggtgttatga gccatattca acgggaaacg tcttgctcta ggccgcgatt aaattccaac
     2101 atggatgctg atttatatgg gtataaatgg gctcgcgata atgtcgggca atcaggtgcg
     2161 acaatctatc gattgtatgg gaagcccgat gcgccagagt tgtttctgaa acatggcaaa
     2221 ggtagcgttg ccaatgatgt tacagatgag atggtcagac taaactggct gacggaattt
     2281 atgcctcttc cgaccatcaa gcattttatc cgtactcctg atgatgcatg gttactcacc
     2341 actgcgatcc ccgggaaaac agcattccag gtattagaag aatatcctga ttcaggtgaa
     2401 aatattgttg atgcgctggc agtgttcctg cgccggttgc attcgattcc tgtttgtaat
     2461 tgtcctttta acagcgatcg cgtatttcgt ctcgctcagg cgcaatcacg aatgaataac
     2521 ggtttggttg atgcgagtga ttttgatgac gagcgtaatg gctggcctgt tgaacaagtc
     2581 tggaaagaaa tgcataaact tttgccattc tcaccggatt cagtcgtcac tcatggtgat
     2641 ttctcacttg ataaccttat ttttgacgag gggaaattaa taggttgtat tgatgttgga
     2701 cgagtcggaa tcgcagaccg ataccaggat cttgccatcc tatggaactg cctcggtgag
     2761 ttttctcctt cattacagaa acggcttttt caaaaatatg gtattgataa tcctgatatg
     2821 aataaattgc agtttcattt gatgctcgat gagtttttct aagaattaat tcatgagcgg
     2881 atacatattt gaatgtattt agaaaaataa acaaataggg gttccgcgca catttccccg
     2941 aaaagtgcca cctgaaattg taaacgttaa tattttgtta aaattcgcgt taaatttttg
     3001 ttaaatcagc tcatttttta accaataggc cgaaatcggc aaaatccctt ataaatcaaa
     3061 agaatagacc gagatagggt tgagtgttgt tccagtttgg aacaagagtc cactattaaa
     3121 gaacgtggac tccaacgtca aagggcgaaa aaccgtctat cagggcgatg gcccactacg
     3181 tgaaccatca ccctaatcaa gttttttggg gtcgaggtgc cgtaaagcac taaatcggaa
     3241 ccctaaaggg agcccccgat ttagagcttg acggggaaag ccggcgaacg tggcgagaaa
     3301 ggaagggaag aaagcgaaag gagcgggcgc tagggcgctg gcaagtgtag cggtcacgct
     3361 gcgcgtaacc accacacccg ccgcgcttaa tgcgccgcta cagggcgcgt cccattcgcc
     3421 aatccggata tagttcctcc tttcagcaaa aaacccctca agacccgttt agaggcccca
     3481 aggggttatg ctagttattg ctcagcggtg gcagcagcca actcagcttc ctttcgggct
     3541 ttgttagcag ccggatctta ttccttgcgc accacctcct cctcctcctg gcgcttctcg
     3601 tagtgagcaa agtcgtcgaa gatggaggtt gtgtgcttgt aggaactgat aattttcagc
     3661 acttgcttgg ctttttctaa gggcacctcc tgggtgtccc gggagtttgt gaccggtttg
     3721 ttgtcgttat tctccagcct gatgtgccgg agctggttat tgggtacatc cttaacaaaa
     3781 atccactgga catcaaactt ccccttccac ttgtcctgag accagacccc ggcactggtg
     3841 ccgtagtcca cgggggactt catctcggcc accccacaaa aatgcccact cccattgacg
     3901 ctgaagagca ggtagacggg ccccttgctg ctcatgcagc ggaaggcgct gtccaggcgc
     3961 ttgttgccgt gctctgtgct acaccagatg gagtacttaa tggagcggtg gatgtcgtcc
     4021 tcagagtagc tcttgatgat gaacacacgc ccgcttttca gattccactc aaactctttc
     4081 gggttgtagc tgtgaccgct tccaccatga tgatgatgat ggtgcatatg tatatctcct
     4141 tcttaaagtt aaacaaaatt atttctagag gggaattgtt atccgctcac aattccccta
     4201 tagtgagtcg tattaatttc gcgggatcga gatcgatctc gatcctctac gccggacgca
     4261 tcgtggccgg catcaccggc gccacaggtg cggttgctgg cgcctatatc gccgacatca
     4321 ccgatgggga agatcgggct cgccacttcg ggctcatgag cgcttgtttc ggcgtgggta
     4381 tggtggcagg ccccgtggcc gggggactgt tgggcgccat ctccttgcat gcaccattcc
     4441 ttgcggcggc ggtgctcaac ggcctcaacc tactactggg ctgcttccta atgcaggagt
     4501 cgcataaggg agagcgtcga gatcccggac accatcgaat ggcgcaaaac ctttcgcggt
     4561 atggcatgat agcgcccgga agagagtcaa ttcagggtgg tgaatgtgaa accagtaacg
     4621 ttatacgatg tcgcagagta tgccggtgtc tcttatcaga ccgtttcccg cgtggtgaac
     4681 caggccagcc acgtttctgc gaaaacgcgg gaaaaagtgg aagcggcgat ggcggagctg
     4741 aattacattc ccaaccgcgt ggcacaacaa ctggcgggca aacagtcgtt gctgattggc
     4801 gttgccacct ccagtctggc cctgcacgcg ccgtcgcaaa ttgtcgcggc gattaaatct
     4861 cgcgccgatc aactgggtgc cagcgtggtg gtgtcgatgg tagaacgaag cggcgtcgaa
     4921 gcctgtaaag cggcggtgca caatcttctc gcgcaacgcg tcagtgggct gatcattaac
     4981 tatccgctgg atgaccagga tgccattgct gtggaagctg cctgcactaa tgttccggcg
     5041 ttatttcttg atgtctctga ccagacaccc atcaacagta ttattttctc ccatgaagac
     5101 ggtacgcgac tgggcgtgga gcatctggtc gcattgggtc accagcaaat cgcgctgtta
     5161 gcgggcccat taagttctgt ctcggcgcgt ctgcgtctgg ctggctggca taaatatctc
     5221 actcgcaatc aaattcagcc gatagcggaa cgggaaggcg actggagtgc catgtccggt
     5281 tttcaacaaa ccatgcaaat gctgaatgag ggcatcgttc ccactgcgat gctggttgcc
     5341 aacgatcaga tggcgctggg cgcaatgcgc gccattaccg agtccgggct gcgcgttggt
     5401 gcggacatct cggtagtggg atacgacgat accgaagaca gctcatgtta tatcccgccg
     5461 ttaaccacca tcaaacagga ttttcgcctg ctggggcaaa ccagcgtgga ccgcttgctg
     5521 caactctctc agggccaggc ggtgaagggc aatcagctgt tgcccgtctc actggtgaaa
     5581 agaaaaacca ccctggcgcc caatacgcaa accgcctctc cccgcgcgtt ggccgattca
     5641 ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc gcaacgcaat
     5701 taatgtaagt tagctcactc attaggcacc gggatctcga ccgatgccct tgagagcctt
     5761 caacccagtc agctccttcc g
//
