LOCUS       Exported                4850 bp ds-DNA     circular UNA 02-MAY-2024
DEFINITION  Rep101 Ts - IS621 WT Donor (22bp) with Bba_R0040-KanR Cargo and 
            Rep101 Temp Sensitive.
ACCESSION   <unknown
VERSION     .
KEYWORDS    .
SOURCE      natural DNA sequence
  ORGANISM  unspecified
REFERENCE   1  (bases 1 to 4850)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jun 13, 2024 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4850
                     /organism="unspecified"
                     /mol_type="genomic DNA"
     rep_origin      1..223
     regulatory      complement(960..1092)
                     /standard_name="BBa_B0015 Terminator"
                     /label=BBa_B0015 Terminator
                     /note="Geneious type: terminator"
     CDS             complement(1102..1917)
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /standard_name="KanR"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /translation="MSHIQRETSCSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYG
                     KPDAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGK
                     TAFQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDA
                     SDFDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGI
                     ADRYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     misc_feature    complement(1918..1942)
                     /standard_name="RBS"
                     /label=RBS
                     /note="Geneious type: RBS"
     regulatory      complement(1943..1996)
                     /standard_name="Bba_R0040 Promoter"
                     /label=Bba_R0040 Promoter
                     /note="Geneious type: promoter"
     misc_feature    2029..2038
                     /standard_name="IS621 RE"
                     /label=IS621 RE
                     /note="Geneious type: RE"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2023.0.3 'Annotate
                     from Database' based on nucleotide similarity"
     misc_feature    2032..2045
                     /standard_name="IS621 WT Donor"
                     /label=IS621 WT Donor
                     /note="Geneious type: Donor"
                     /note="Derived using Geneious Prime 2023.0.3 'Annotate
                     from Database' based on nucleotide similarity"
     misc_feature    2032..2040
                     /standard_name="LD"
                     /label=LD
                     /note="Geneious type: LD"
     misc_feature    complement(2039..2045)
                     /standard_name="RD"
                     /label=RD
                     /note="Geneious type: RD"
     misc_feature    2039..2040
                     /standard_name="Core"
                     /label=Core
                     /note="Geneious type: Core"
     misc_feature    2041..2050
                     /standard_name="IS621 LE"
                     /label=IS621 LE
                     /note="Geneious type: LE"
                     /note="Derived using Geneious Prime 2022.0.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /note="Derived using Geneious Prime 2023.0.3 'Annotate
                     from Database' based on nucleotide similarity"
     regulatory      2371..2448
                     /gene="lacI"
                     /standard_name="lacI promoter"
                     /label=lacI promoter
                     /note="Geneious type: promoter"
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
     CDS             2449..3531
                     /codon_start=1
                     /gene="lacI"
                     /standard_name="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)."
                     /note="Derived using Geneious Prime 2021.2.2 'Annotate
                     from Database' based on nucleotide similarity"
                     /translation="VKPVTLYDVAEYAGVSYQTVSRVVNQASHVSAKTREKVEAAMAEL
                     NYIPNRVAQQLAGKQSLLIGVATSSLALHAPSQIVAAIKSRADQLGASVVVSMVERSGV
                     EACKAAVHNLLAQRVSGLIINYPLDDQDAIAVEAACTNVPALFLDVSDQTPINSIIFSH
                     EDGTRLGVEHLVALGHQQIALLAGPLSSVSARLRLAGWHKYLTRNQIQPIAEREGDWSA
                     MSGFQQTMQMLNEGIVPTAMLVANDQMALGAMRAITESGLRVGADISVVGYDDTEDSSC
                     YIPPLTTIKQDFRLLGQTSVDRLLQLSQGQAVKGNQLLPVSLVKRKTTLAPNTQTASPR
                     ALADSLMQLARQVSRLESGQ"
     CDS             complement(3853..4803)
                     /standard_name="Rep101(Ts)"
                     /label=Rep101(Ts)
                     /note="pLannotate"
ORIGIN
        1 gggctagtca atgataatta ctagtccttt tcctttgagt tgtgggtatc tgtaaattct
       61 gctagacctt tgctggaaaa cttgtaaatt ctgctagacc ctctgtaaat tccgctagac
      121 ctttgtgtgt tttttttgtt tatattcaag tggttataat ttatagaata aagaaagaat
      181 aaaaaaagat aaaaagaata gatcccagcc ctgtgtataa ctcactactt tagtcagttc
      241 cgcagtatta caaaaggatg tcgcaaacgc tgtttgctcc tctacaaaac agaccttaaa
      301 accctaaagg cttaagtagc accctcgcaa gctcgggcaa atcgctgaat attccttttg
      361 tctccgacca tcaggcacct gagtcgctgt ctttttcgtg acattcagtt cgctgcgctc
      421 acggctctgg cagtgaatgg gggtaaatgg cactacaggc gccttttatg gattcatgca
      481 aggaaactac ccataataca agaaaagccc gtcacgggct tctcagggcg ttttatggcg
      541 ggtctgctat gtggtgctat ctgacttttt gctgttcagc agttcctgcc ctctgatttt
      601 ccagtctgac cacttcggat tatcccgtga caggtcattc agactggcta atgcacccag
      661 taaggcagcg gtatcatcaa ccaggtggca cttttcgggg aaatgtgttg gggtcgaggt
      721 gccgtaaagc actaaatcgg aaccctaaag ggagcccccg atttagagct tgacggggaa
      781 agccggcgaa cgtggcgaga aaggaaggga agaaagcgaa aggagcgggc gctagggcgc
      841 tggcaagtgt agcggtcacg ctgcgcgtaa ccaccacacc cgccgcgctt aatgcgccgc
      901 tacagggcgc gtcccattcg ccaatccgga tatagttcct ccttttggtg gtgctcgagt
      961 ctatataaac gcagaaaggc ccacccgaag gtgagccagt gtgactctag tagagagcgt
     1021 tcaccgacaa acaacagata aaacgaaagg cccagtcttt cgactgagcc tttcgtttta
     1081 tttgatgcct ggagatcctt attagaaaaa ctcatcgagc atcaaatgaa actgcaattt
     1141 attcatatca ggattatcaa taccatattt ttgaaaaagc cgtttctgta atgaaggaga
     1201 aaactcaccg aggcagttcc ataggatggc aagatcctgg tatcggtctg cgattccgac
     1261 tcgtccaaca tcaatacaac ctattaattt cccctcgtca aaaataaggt tatcaagtga
     1321 gaaatcacca tgagtgacga ctgaatccgg tgagaatggc aaaagtttat gcatttcttt
     1381 ccagacttgt tcaacaggcc agccattacg ctcgtcatca aaatcactcg catcaaccaa
     1441 accgttattc attcgtgatt gcgcctgagc gagacgaaat acgcgatcgc tgttaaaagg
     1501 acaattacaa acaggaatcg aatgcaaccg gcgcaggaac actgccagcg catcaacaat
     1561 attttcacct gaatcaggat attcttctaa tacctggaat gctgttttcc cggggatcgc
     1621 agtggtgagt aaccatgcat catcaggagt acggataaaa tgcttgatgg tcggaagagg
     1681 cataaattcc gtcagccagt ttagtctgac catctcatct gtaacatcat tggcaacgct
     1741 acctttgcca tgtttcagaa acaactctgg cgcatcgggc ttcccataca atcgatagat
     1801 tgtcgcacct gattgcccga cattatcgcg agcccattta tacccatata aatcagcatc
     1861 catgttggaa tttaatcgcg gcctagagca agacgtttcc cgttgaatat ggctcataga
     1921 tcatttctcc tctttcagat ccgtgctcag tatctctatc actgataggg atgtcaatct
     1981 ctatcactga tagggaggac ctcctaaata gttagtttga cgcacataat aacagtatct
     2041 tgtattatcc ggatgtgccc atggtatatc tccttcttat ctcgatcctc tacgccggac
     2101 gcatcgtggc cggcatcacc ggcgccacag gtgcggttgc tggcgcctat atcgccgaca
     2161 tcaccgatgg ggaagatcgg gctcgccact tcgggctcat gagcgcttgt ttcggcgtgg
     2221 gtatggtggc aggccccgtg gccgggggac tgttgggcgc catctccttg catgcaccat
     2281 tccttgcggc ggcggtgctc aacggcctca acctactact gggctgcttc ctaatgcagg
     2341 agtcgcataa gggagagcgt cgagatcccg gacaccatcg aatggcgcaa aacctttcgc
     2401 ggtatggcat gatagcgccc ggaagagagt caattcaggg tggtgaatgt gaaaccagta
     2461 acgttatacg atgtcgcaga gtatgccggt gtctcttatc agaccgtttc ccgcgtggtg
     2521 aaccaggcca gccacgtttc tgcgaaaacg cgggaaaaag tggaagcggc gatggcggag
     2581 ctgaattaca ttcccaaccg cgtggcacaa caactggcgg gcaaacagtc gttgctgatt
     2641 ggcgttgcca cctccagtct ggccctgcac gcgccgtcgc aaattgtcgc ggcgattaaa
     2701 tctcgcgccg atcaactggg tgccagcgtg gtggtgtcga tggtagaacg aagcggcgtc
     2761 gaagcctgta aagcggcggt gcacaatctt ctcgcgcaac gcgtcagtgg gctgatcatt
     2821 aactatccgc tggatgacca ggatgccatt gctgtggaag ctgcctgcac taatgttccg
     2881 gcgttatttc ttgatgtctc tgaccagaca cccatcaaca gtattatttt ctcccatgaa
     2941 gacggtacgc gactgggcgt ggagcatctg gtcgcattgg gtcaccagca aatcgcgctg
     3001 ttagcgggcc cattaagttc tgtctcggcg cgtctgcgtc tggctggctg gcataaatat
     3061 ctcactcgca atcaaattca gccgatagcg gaacgggaag gcgactggag tgccatgtcc
     3121 ggttttcaac aaaccatgca aatgctgaat gagggcatcg ttcccactgc gatgctggtt
     3181 gccaacgatc agatggcgct gggcgcaatg cgcgccatta ccgagtccgg gctgcgcgtt
     3241 ggtgcggata tctcggtagt gggatacgac gataccgaag acagctcatg ttatatcccg
     3301 ccgttaacca ccatcaaaca ggattttcgc ctgctggggc aaaccagcgt ggaccgcttg
     3361 ctgcaactct ctcagggcca ggcggtgaag ggcaatcagc tgttgcccgt ctcactggtg
     3421 aaaagaaaaa ccaccctggc gcccaatacg caaaccgcct ctccccgcgc gttggccgat
     3481 tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc
     3541 aattaatgta agttagctca ctcattaggc accgggatct cgaccgatgc ccttgagagc
     3601 cttcaaccca gtcagctcct tggataagag acggctcaac gcccgtctca cagcgcaata
     3661 tcaatttatg ggtgtgataa ttatcaattt atgggtgtaa ttatcatttt atggttgtat
     3721 caacaaggag gactttggca ggtgcctgca ggtatggaca gttttccctt tgatatgtaa
     3781 cggtgaacag ttgttctact tttgtttgtt agtcttgatg cttcactgat agatacaaga
     3841 gccataagaa cctcagatcc ttccgtattt agccagtatg ttctctagtg tggttcgttg
     3901 tttttgcgtg agccatgaga acgaaccatt gagatcatac ttactttgca tgtcactcaa
     3961 aaattttgcc tcaaaactgg tgagctgaat ttttgcagtt aaagcatcgt gtagtgtttt
     4021 tcttagtccg ttatgtaggt aggaatctga tgtaatggtt gttggtattt tgtcaccatt
     4081 catttttatc tggttgttct caagttcggt tacgagatcc atttgtctat ctagttcaac
     4141 ttggaaaatc aacgtatcag tcgggcggcc tcgcttatca accaccaatt tcatattgct
     4201 gtaagtgttt aaatctttac ttattggttt caaaacccat tggttaagcc ttttaaactc
     4261 atggtagtta ttttcaagca ttaacatgaa cttaaattca tcaaggctaa tctctatatt
     4321 tgccttgtga gttttctttt gtgttagttc ttttaataac cactcataaa tcctcataga
     4381 gtatttgttt tcaaaagact taacatgttc cagattatat tttatgaatt tttttaactg
     4441 gaaaagataa ggcaatatct cttcactaaa aactaattct aatttttcgc ttgagaactt
     4501 ggcatagttt gtccactgga aaatctcaaa gcctttaacc aaaggattcc tgatttccac
     4561 agttctcgtc atcagctctc tggttgcttt agctaataca ccataagcat tttccctact
     4621 gatgttcatc atctgaacgt attggttata agtgaacgat accgtccgtt ctttccttgt
     4681 agggttttca atcgtggggt tgagtagtgc cacacagcat aaaattagct tggtttcatg
     4741 ctccgttaag tcatagcgac taatcgctag ttcatttgct ttgaaaacaa ctaattcaga
     4801 catacatctc aattggtcta ggtgatttta atcactatac caattgagat
//
