LOCUS       LRTX_ctrl        7643 bp DNA     circular SYN 28-DEC-2023
DEFINITION  LRTX8  (7643 bp).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7643)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 28, 2023 from SnapGene Viewer 7.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7643
                     /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"
     regulatory      3316..3325
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     CDS             3325..3354
                     /codon_start=1
                     /product="Myc (human c-Myc proto-oncogene) epitope tag"
                     /label=Myc
                     /translation="EQKLISEEDL"
     CDS             3355..5001
                     /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    5068..5189
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     polyA_signal    5285..5396
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     CDS             complement(5538..6137)
                     /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(6142..6175)
                     /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    6387..6435
                     /label=HSV TK poly(A) signal
                     /note="herpes simplex virus thymidine kinase 
                     polyadenylation signal (Cole and Stacy, 1985)"
     promoter        6508..6612
                     /gene="bla"
                     /label=AmpR promoter
     CDS             6613..7473
                     /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 ataccggtcg ccaccgccac catggaacaa aagctgattt ctgaagaaga cttggaagac
     3361 gccaaaaaca taaagaaagg cccggcgcca ttctatccgc tggaagatgg aaccgctgga
     3421 gagcaactgc ataaggctat gaagagatac gccctggttc ctggaataat tgcttttaca
     3481 gatgcacata tcgaggtgga catcacttac gctgagtact tcgaaatgtc cgttcggttg
     3541 gcagaagcta tgaaacgata tgggctgaat acaaatcaca gaatcgtcgt atgcagtgaa
     3601 aactctcttc aattctttat gccggtgttg ggcgcgttat ttatcggagt tgcagttgcg
     3661 cccgcgaacg acatttataa tgaacgtgaa ttgctcaaca gtatgggcat ttcgcagcct
     3721 accgtggtgt tcgtttccaa aaaggggttg caaaaaattt tgaacgtgca aaaaaagctc
     3781 ccaatcatcc aaaaaattat tatcatggat tctaaaacgg attaccaggg atttcagtcg
     3841 atgtacacgt tcgtcacatc tcatctacct cccggtttta atgaatacga ttttgtgcca
     3901 gagtccttcg atagggacaa gataattgca ctgatcatga actcctctgg atctactggt
     3961 ctgcctaaag gtgtcgctct gcctcataga actgcctgcg tgagattctc gcatgccaga
     4021 gatcctattt ttggcaatca aatcattccg gatactgcga ttttaagtgt tgttccattc
     4081 catcacggtt ttggaatgtt tactacactc ggatatttga tatgtggatt tcgagtcgtc
     4141 ttaatgtata gatttgaaga agagctgttt ctgaggagcc ttcaggatta caagattcaa
     4201 agtgcgctgc tggtgccaac cctattctcc ttcttcgcca aaagcactct gattgacaaa
     4261 tacgatttat ctaatttaca cgaaattgct tctggtggcg ctcccctctc taaggaagtc
     4321 ggggaagcgg ttgccaagag gttccatctg ccaggtatca ggcaaggata tgggctcact
     4381 gagactacat cagctattct gattacaccc gagggggatg ataaaccggg cgcggtcggt
     4441 aaagttgttc cattttttga agcgaaggtt gtggatctgg ataccgggaa aacgctgggc
     4501 gttaatcaaa gaggcgaact gtgtgtgaga ggtcctatga ttatgtccgg ttatgtaaac
     4561 aatccggaag cgaccaacgc cttgattgac aaggatggat ggctacattc tggagacata
     4621 gcttactggg acgaagacga acacttcttc atcgttgacc gcctgaagtc tctgattaag
     4681 tacaaaggct atcaggtggc tcccgctgaa ttggaatcca tcttgctcca acaccccaac
     4741 atcttcgacg caggtgtcgc aggtcttccc gacgatgacg ccggtgaact tcccgccgcc
     4801 gttgttgttt tggagcacgg aaagacgatg acggaaaaag agatcgtgga ttacgtcgcc
     4861 agtcaagtaa caaccgcgaa aaagttgcgc ggaggagttg tgtttgtgga cgaagtaccg
     4921 aaaggtctta ccggaaaact cgacgcaaga aaaatcagag agatcctcat aaaggccaag
     4981 aagggcggaa agatcgccgt gctcgagggt ggttctggtg gttctgccta aggtggttct
     5041 ggtggttctg aattaattgt tgttgttaac ttgtttattg cagcttataa tggttacaaa
     5101 taaagcaata gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt
     5161 ggtttgtcca aactcatcaa tgtatcttaa cgcgaaacta gtgctattgt cttcccaatc
     5221 ctcccccttg ctgtcctgcc ccaccccacc ccccagaata gaatgacacc tactcagaca
     5281 atgcgatgca atttcctcat tttattagga aaggacagtg ggagtggcac cttccagggt
     5341 caaggaaggc acgggggagg ggcaaacaac agatggctgg caactagaag gcacagtcga
     5401 ggctgatcag cgagctctag agtcggtggg cctcgggggc gggtgcgggg tcggcggggc
     5461 cgcccgggtc ggcttcggtc ggagccatgg ggtcgtgcgc tcctttcggt cgggcgctgc
     5521 gggtcgtggg gcgggcgtca ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct
     5581 tcgggcacct cgacgtcggc ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg
     5641 cggggcgcgg aggtctccag gaaggcgggc accccggcgc gctcggccgc ctccactccg
     5701 gggagcacga cggcgctgcc cagacccttg ccctggtggt cgggcgacac gccgacggtg
     5761 gccaggaacc acgcgggctc cttgggccgg tgcggcgcca ggaggccttc catctgttgc
     5821 tgcgcggcca gccgggaacc gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc
     5881 gcccccgctt cgacgctctc cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg
     5941 acccacacct tgccgatgtc gagcccgacg cgcgtgagga agagttcttg cagctcggtg
     6001 acccgctcga tgtggcggtc cgggtcgacg gtgtggcgcg tggcggggta gtcggcgaac
     6061 gcggcggcga gggtgcgtac ggcccggggg acgtcgtcgc gggtggcgag gcgcaccgtg
     6121 ggcttgtact cggtcatggt gataacttcg tataggatac tttatacgaa gttatttatt
     6181 ctcgaatggg ggaccccgtc cctaacccac ggggccagtg gctatggcag ggcctgccgc
     6241 cccgacgttg gctgcgagcc ctgggccttc acccgaactt ggggggtggg gtggggaaaa
     6301 ggaagaaacg cgggcgtatt ggccccaatg gggtctcggt ggggtatcga cagagtgcca
     6361 gccctgggac cgaaccccgc gtttatgaac aaacgaccca acacccgtgc gttttattct
     6421 gtctttttat tgccgtcata gcgcgggttc cttccggtat tgtctccttc cgtgtttatt
     6481 aagcttggca cttttcgggg aaatgtgcgc ggaaccccta tttgtttatt tttctaaata
     6541 cattcaaata tgtatccgct catgagacaa taaccctgat aaatgcttca ataatattga
     6601 aaaaggaaga gtatgagtat tcaacatttc cgtgtcgccc ttattccctt ttttgcggca
     6661 ttttgccttc ctgtttttgc tcacccagaa acgctggtga aagtaaaaga tgctgaagat
     6721 cagttgggtg cacgagtggg ttacatcgaa ctggatctca acagcggtaa gatccttgag
     6781 agttttcgcc ccgaagaacg ttttccaatg atgagcactt ttaaagttct gctatgtggc
     6841 gcggtattat cccgtattga cgccgggcaa gagcaactcg gtcgccgcat acactattct
     6901 cagaatgact tggttgagta ctcaccagtc acagaaaagc atcttacgga tggcatgaca
     6961 gtaagagaat tatgcagtgc tgccataacc atgagtgata acactgcggc caacttactt
     7021 ctgacaacga tcggaggacc gaaggagcta accgcttttt tgcacaacat gggggatcat
     7081 gtaactcgcc ttgatcgttg ggaaccggag ctgaatgaag ccataccaaa cgacgagcgt
     7141 gacaccacga tgcctgtagc aatggcaaca acgttgcgca aactattaac tggcgaacta
     7201 cttactctag cttcccggca acaattaata gactggatgg aggcggataa agttgcagga
     7261 ccacttctgc gctcggccct tccggctggc tggtttattg ctgataaatc tggagccggt
     7321 gagcgtgggt ctcgcggtat cattgcagca ctggggccag atggtaagcc ctcccgtatc
     7381 gtagttatct acacgacggg gagtcaggca actatggatg aacgaaatag acagatcgct
     7441 gagataggtg cctcactgat taagcattgg taactgtcag accaagttta ctcatatata
     7501 ctttagattg atttaaaact tcatttttaa tttaaaagga tctaggtgaa gatccttttt
     7561 gataatctca tgaccaaaat cccttaacgt gagttttcgt tccactgagc gtcagacccc
     7621 gtagaaaaga tcaaaggatc ttc
//
