LOCUS       LiRTX_ctrl        7937 bp DNA     circular SYN 28-DEC-2023
DEFINITION  LiRTX8  (7937 bp).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7937)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 28, 2023 from SnapGene Viewer 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7937
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    877..898
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        913..943
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    951..967
                     /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     975..991
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        1012..1030
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     polyA_signal    1049..1170
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     protein_bind    complement(1183..1216)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     polyA_signal    1448..1503
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     CDS             complement(1645..2577)
                     /codon_start=1
                     /product="luciferase from the anthozoan coelenterate 
                     Renilla reniformis (sea pansy)"
                     /label=Rluc
                     /translation="TSKVYDPEQRKRMITGPQWWARCKQMNVLDSFINYYDSEKHAENA
                     VIFLHGNAASSYLWRHVVPHIEPVARCIIPDLIGMGKSGKSGNGSYRLLDHYKYLTAWF
                     ELLNLPKKIIFVGHDWGACLAFHYSYEHQDKIKAIVHAESVVDVIESWDEWPDIEEDIA
                     LIKSEEGEKMVLENNFFVETMLPSKIMRKLEPEEFAAYLEPFKEKGEVRRPTLSWPREI
                     PLVKGGKPDVVQIVRNYNAYLRASDDLPKMFIESDPGFFSNAIVEGAKKFPNTEFVKVK
                     GLHFSQEDAPDEMGKYIKSFVERVLKNEQ"
     CDS             complement(2578..2595)
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     regulatory      2601..2610
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     enhancer        2775..3078
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3079..3282
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     ncRNA           3409..3435
                     /label=5' miR-155
                     /note="sequence upstream of the precursor of mouse miR-155 
                     microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     ncRNA           3461..3502
                     /label=3' miR-155
                     /note="sequence downstream of the precursor of mouse 
                     miR-155 microRNA (Uva et al., 2013)"
                     /ncRNA_class="miRNA"
     regulatory      3610..3619
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3619..3648
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3649..5295
                     /codon_start=1
                     /gene="luc+"
                     /product="firefly luciferase"
                     /label=luciferase
                     /note="enhanced luc+ version of the luciferase gene"
                     /translation="EDAKNIKKGPAPFYPLEDGTAGEQLHKAMKRYALVPGIIAFTDAH
                     IEVDITYAEYFEMSVRLAEAMKRYGLNTNHRIVVCSENSLQFFMPVLGALFIGVAVAPA
                     NDIYNERELLNSMGISQPTVVFVSKKGLQKILNVQKKLPIIQKIIIMDSKTDYQGFQSM
                     YTFVTSHLPPGFNEYDFVPESFDRDKIIALIMNSSGSTGLPKGVALPHRTACVRFSHAR
                     DPIFGNQIIPDTAILSVVPFHHGFGMFTTLGYLICGFRVVLMYRFEEELFLRSLQDYKI
                     QSALLVPTLFSFFAKSTLIDKYDLSNLHEIASGGAPLSKEVGEAVAKRFHLPGIRQGYG
                     LTETTSAILITPEGDDKPGAVGKVVPFFEAKVVDLDTGKTLGVNQRGELCVRGPMIMSG
                     YVNNPEATNALIDKDGWLHSGDIAYWDEDEHFFIVDRLKSLIKYKGYQVAPAELESILL
                     QHPNIFDAGVAGLPDDDAGELPAAVVVLEHGKTMTEKEIVDYVASQVTTAKKLRGGVVF
                     VDEVPKGLTGKLDARKIREILIKAKKGGKIAV"
     polyA_signal    5362..5483
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    5579..5690
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(5832..6431)
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /product="puromycin N-acetyltransferase"
                     /label=PuroR
                     /note="confers resistance to puromycin"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     protein_bind    complement(6436..6469)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (AAGTATCC) (Shaw et al., 2021). lox2272 sites are 
                     compatible with each other, but incompatible with loxP or 
                     loxN sites (Lee and Saito, 1988)."
     polyA_signal    6681..6729
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6802..6906
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6907..7767
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcggaag agcgcccaat
      781 acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc acgacaggtt
      841 tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc tcactcatta
      901 ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa ttgtgagcgg
      961 ataacaattt cacacaggaa acagctatga ccatgattac gccaagcgcg caattaaccc
     1021 tcactaaagg gaacaaaagc tgggtaccta agatacattg atgagtttgg acaaaccaca
     1081 actagaatgc agtgaaaaaa atgctttatt tgtgaaattt gtgatgctat tgctttattt
     1141 gtaaccatta taagctgcaa taaacaagtt gacggtatcg taataacttc gtatagcata
     1201 cattatacga agttatatgg atccatatgc tagctcgacg ctcaggtcga gggatcttca
     1261 taagagaaga gggacagcta tgactgggag taaggaatgg acagcagggg gctgtttcat
     1321 atactgatga cctctttata gccacctttg ttcatggcag ccagcatatg gcatatgttg
     1381 ccaaactcta aaccaaatac tcattctgat gttttaaatg atttgccctc ccatatgtcc
     1441 ttccgagtga gagacacaaa aaattccaac acactattgc aatgaaaata aatttccttt
     1501 attagccaga agtcagatgc tcaaggggct tcatgatgtc cccataattt ttggcagagg
     1561 gaaaaagatc tcagtggtat ttgtgagcca gggcattggc cacaccagcc accaccttct
     1621 gataggcagc ctgcacctga gtgtttattg ttcatttttg agaactcgct caacgaacga
     1681 tttgatatat tttcccattt catcaggtgc atcttcttgc gaaaaatgaa gaccttttac
     1741 tttgacaaat tcagtattag gaaacttctt ggcaccttca acaatagcat tggaaaagaa
     1801 tcctgggtcc gattcaataa acatttttgg taaatcatca cttgcacgta gataagcatt
     1861 ataattccta acaatttgta caacgtcagg tttaccacct tttactaacg ggatttcacg
     1921 aggccatgat aatgttggac gacgaacttc acctttctct ttgaatggtt caagatatgc
     1981 tgcaaattct tctggttcta actttctcat gatttttgat ggcaacatgg tttccacgaa
     2041 gaagttattc tccaaaacca ttttttctcc ttcttcagat ttgatcaacg caatatcttc
     2101 ttcaatatca ggccattcat cccatgattc aatcacatct actacacttt cagcgtgaac
     2161 tattgctttg atcttatctt gatgctcata gctataatga aatgccaaac aagcacccca
     2221 atcatggccg acaaaaatga tcttctttgg taaattaaga agttcaaacc atgcagtaag
     2281 atatttgtaa tgatcaagta acctataaga accattacca gatttgcctg atttgcccat
     2341 accaataagg tctggtataa tacaccgcgc tactggctca atatgtggca caacatgtcg
     2401 ccataaataa gaagaggccg cgttaccatg taaaaaaata acagcatttt ctgcatgttt
     2461 ttctgaatca taataattaa taaatgaatc aagaacattc atttgtttac atctggccca
     2521 ccactgcgga ccagttatca tccgtttcct ttgttctgga tcataaactt tcgaagtgtg
     2581 gtgatggtga tgatgatcca ccatggtggc ggtggcgacc acgatcctca tcctgtctct
     2641 gcgatctgac ggttcactaa acgagctctg cttatatagg cctcccaccg tacacgccac
     2701 ctcgacatac tcagtagtta ttaatagtaa tcaattacgg ggtcattagt tcatagcgag
     2761 gctgcagtct accgcgttac ataacttacg gtaaatggcc cgcctggctg accgcccaac
     2821 gacccccgcc cattgacgtc aataatgacg tatgttccca tagtaacgcc aatagggact
     2881 ttccattgac gtcaatgggt ggagtattta cggtaaactg cccacttggc agtacatcaa
     2941 gtgtatcata tgccaagtac gccccctatt gacgtcaatg acggtaaatg gcccgcctgg
     3001 cattatgccc agtacatgac cttatgggac tttcctactt ggcagtacat ctacgtatta
     3061 gtcatcgcta ttaccatggt gatgcggttt tggcagtaca tcaatgggcg tggatagcgg
     3121 tttgactcac ggggatttcc aagtctccac cccattgacg tcaatgggag tttgttttgg
     3181 caccaaaatc aacgggactt tccaaaatgt cgtaacaact ccgccccatt gacgcaaatg
     3241 ggcggtaggc gtgtacggtg ggaggtctat ataagcagag ctggtttagt gaaccgtcag
     3301 ataccggcag gtaagtatca aggttacaag acaggtttaa ggagaccaat agaaactggg
     3361 cttgtcgaga cagagaagac tcttgcgttt cggtcgactt ctagagctct ggaggcttgc
     3421 tgaaggctgt atgctagaga cgtacagatg cgtctcacag gacacaaggc ctgttactag
     3481 cactcacatg gaacaaatgg ccgaattcgg cgctagctgc tgatatcgca tacgcgtgga
     3541 ccagataggc acctattggt cttactgaca tccactttgc ctttctctcc acaggtgccg
     3601 gtcgccaccg ccaccatgga acaaaagctg atttctgaag aagacttgga agacgccaaa
     3661 aacataaaga aaggcccggc gccattctat ccgctggaag atggaaccgc tggagagcaa
     3721 ctgcataagg ctatgaagag atacgccctg gttcctggaa taattgcttt tacagatgca
     3781 catatcgagg tggacatcac ttacgctgag tacttcgaaa tgtccgttcg gttggcagaa
     3841 gctatgaaac gatatgggct gaatacaaat cacagaatcg tcgtatgcag tgaaaactct
     3901 cttcaattct ttatgccggt gttgggcgcg ttatttatcg gagttgcagt tgcgcccgcg
     3961 aacgacattt ataatgaacg tgaattgctc aacagtatgg gcatttcgca gcctaccgtg
     4021 gtgttcgttt ccaaaaaggg gttgcaaaaa attttgaacg tgcaaaaaaa gctcccaatc
     4081 atccaaaaaa ttattatcat ggattctaaa acggattacc agggatttca gtcgatgtac
     4141 acgttcgtca catctcatct acctcccggt tttaatgaat acgattttgt gccagagtcc
     4201 ttcgataggg acaagataat tgcactgatc atgaactcct ctggatctac tggtctgcct
     4261 aaaggtgtcg ctctgcctca tagaactgcc tgcgtgagat tctcgcatgc cagagatcct
     4321 atttttggca atcaaatcat tccggatact gcgattttaa gtgttgttcc attccatcac
     4381 ggttttggaa tgtttactac actcggatat ttgatatgtg gatttcgagt cgtcttaatg
     4441 tatagatttg aagaagagct gtttctgagg agccttcagg attacaagat tcaaagtgcg
     4501 ctgctggtgc caaccctatt ctccttcttc gccaaaagca ctctgattga caaatacgat
     4561 ttatctaatt tacacgaaat tgcttctggt ggcgctcccc tctctaagga agtcggggaa
     4621 gcggttgcca agaggttcca tctgccaggt atcaggcaag gatatgggct cactgagact
     4681 acatcagcta ttctgattac acccgagggg gatgataaac cgggcgcggt cggtaaagtt
     4741 gttccatttt ttgaagcgaa ggttgtggat ctggataccg ggaaaacgct gggcgttaat
     4801 caaagaggcg aactgtgtgt gagaggtcct atgattatgt ccggttatgt aaacaatccg
     4861 gaagcgacca acgccttgat tgacaaggat ggatggctac attctggaga catagcttac
     4921 tgggacgaag acgaacactt cttcatcgtt gaccgcctga agtctctgat taagtacaaa
     4981 ggctatcagg tggctcccgc tgaattggaa tccatcttgc tccaacaccc caacatcttc
     5041 gacgcaggtg tcgcaggtct tcccgacgat gacgccggtg aacttcccgc cgccgttgtt
     5101 gttttggagc acggaaagac gatgacggaa aaagagatcg tggattacgt cgccagtcaa
     5161 gtaacaaccg cgaaaaagtt gcgcggagga gttgtgtttg tggacgaagt accgaaaggt
     5221 cttaccggaa aactcgacgc aagaaaaatc agagagatcc tcataaaggc caagaagggc
     5281 ggaaagatcg ccgtgctcga gggtggttct ggtggttctg cctaaggtgg ttctggtggt
     5341 tctgaattaa ttgttgttgt taacttgttt attgcagctt ataatggtta caaataaagc
     5401 aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag ttgtggtttg
     5461 tccaaactca tcaatgtatc ttaacgcgaa actagtgcta ttgtcttccc aatcctcccc
     5521 cttgctgtcc tgccccaccc caccccccag aatagaatga cacctactca gacaatgcga
     5581 tgcaatttcc tcattttatt aggaaaggac agtgggagtg gcaccttcca gggtcaagga
     5641 aggcacgggg gaggggcaaa caacagatgg ctggcaacta gaaggcacag tcgaggctga
     5701 tcagcgagct ctagagtcgg tgggcctcgg gggcgggtgc ggggtcggcg gggccgcccc
     5761 gggtggcttc ggtcggagcc atggggtcgt gcgctccttt cggtcgggcg ctgcgggtcg
     5821 tggggcgggc gtcaggcacc gggcttgcgg gtcatgcacc aggtgcgcgg tccttcgggc
     5881 acctcgacgt cggcggtgac ggtgaagccg agccgctcgt agaaggggag gttgcggggc
     5941 gcggaggtct ccaggaaggc gggcaccccg gcgcgctcgg ccgcctccac tccggggagc
     6001 acgacggcgc tgcccagacc cttgccctgg tggtcgggcg acacgccgac ggtggccagg
     6061 aaccacgcgg gctccttggg ccggtgcggc gccaggaggc cttccatctg ttgctgcgcg
     6121 gccagccggg aaccgctcaa ctcggccatg cgcgggccga tctcggcgaa caccgccccc
     6181 gcttcgacgc tctccggcgt ggtccagacc gccaccgcgg cgccgtcgtc cgcgacccac
     6241 accttgccga tgtcgagccc gacgcgcgtg aggaagagtt cttgcagctc ggtgacccgc
     6301 tcgatgtggc ggtccgggtc gacggtgtgg cgcgtggcgg ggtagtcggc gaacgcggcg
     6361 gcgagggtgc gtacggcccg ggggacgtcg tcgcgggtgg cgaggcgcac cgtgggcttg
     6421 tactcggtca tggtgataac ttcgtatagg atactttata cgaagttatt tattctcgaa
     6481 tgggggaccc cgtccctaac ccacggggcc agtggctatg gcagggcctg ccgccccgac
     6541 gttggctgcg agccctgggc cttcacccga acttgggggg tggggtgggg aaaaggaaga
     6601 aacgcgggcg tattggcccc aatggggtct cggtggggta tcgacagagt gccagccctg
     6661 ggaccgaacc ccgcgtttat gaacaaacga cccaacaccc gtgcgtttta ttctgtcttt
     6721 ttattgccgt catagcgcgg gttccttccg gtattgtctc cttccgtgtt tattaagctt
     6781 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     6841 aatatgtatc cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg
     6901 aagagtatga gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc
     6961 cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg
     7021 ggtgcacgag tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt
     7081 cgccccgaag aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta
     7141 ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat
     7201 gacttggttg agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga
     7261 gaattatgca gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca
     7321 acgatcggag gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact
     7381 cgccttgatc gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc
     7441 acgatgcctg tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact
     7501 ctagcttccc ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt
     7561 ctgcgctcgg cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt
     7621 gggtctcgcg gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt
     7681 atctacacga cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata
     7741 ggtgcctcac tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag
     7801 attgatttaa aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat
     7861 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     7921 aagatcaaag gatcttc
//
