LOCUS       Exported                4703 bp ds-DNA     circular SYN 16-AUG-2022
DEFINITION  .
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4703)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Apr 4, 2023 from SnapGene 4.3.11
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..4703
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     terminator      1..87
                     /gene="Escherichia coli rrnB"
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
     primer_bind     21..40
                     /label=A423 rrnb T1 ter -f
                     /note="color: black; sequence: cgttcaccgacaaacaacag; added:
                     2022-07-25"
     primer_bind     127..146
                     /label=C133 pME 5' con -f
                     /note="color: black; sequence: ggatgttttcccagtcacga; added:
                     2022-07-25"
     primer_bind     136..158
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
                     /note="color: black; sequence: CCCAGTCACGACGTTGTAAAACG"
     primer_bind     150..167
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
                     /note="color: black; sequence: TGTAAAACGACGGCCAGT"
     primer_bind     151..167
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
                     /note="color: #a020f0; direction: RIGHT"
     primer_bind     182..225
                     /label=A1151 CreER-BsaI-ins-f
                     /note="color: black; sequence: 
                     cggtctctAGTCgccaccatgtccaatttactgaccgtacacca; added: 
                     2022-08-09"
     primer_bind     complement(182..221)
                     /label=A1150 BsaI-CreER-vec-r
                     /note="color: black; sequence: 
                     gtacggtcagtaaattggacatggtggcGACTagagaccg; added: 
                     2022-08-09"
     regulatory      194..199
                     /regulatory_class="other"
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /note="color: #a6acb3"
     primer_bind     197..225
                     /label=Cre KpnI -f
                     /note="color: black; sequence: 
                     ccggtaccatgtccaatttactgaccgtacacca"
     CDS             200..1228
                     /codon_start=1
                     /product="site-specific recombinase"
                     /label=Cre
                     /note="Cre is a site-specific recombinase from 
                     bacteriophage P1. Recombination occurs at loxP sequences."
                     /note="color: #993366"
                     /translation="MSNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKML
                     LSVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGL
                     PRPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAF
                     LGIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVE
                     RWISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQ
                     RYLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLE
                     DGD"
     primer_bind     758..777
                     /label=C37 Cre Scr -f
                     /note="color: black; sequence: cgtactgacggtgggagaat; added:
                     2022-08-17"
     primer_bind     complement(857..866)
                     /label=A228 EGFP-N
                     /note="color: black; sequence: cgtcgccgtccagctcgaccag; 
                     added: 2022-02-04"
     primer_bind     complement(937..957)
                     /label=C38 Cre Scr -r
                     /note="color: black; sequence: gtggcagatggcgcggcaaca; 
                     added: 2022-08-17"
     primer_bind     complement(1209..1228)
                     /label=Cre-stop-r
                     /note="color: black; sequence: ctaatcgccatcttccagcaggc"
     CDS             1241..2176
                     /codon_start=1
                     /product="mutated ligand-binding domain of the human 
                     estrogen receptor (Feil et al., 1997)"
                     /label=ERT2
                     /note="color: #993366"
                     /translation="AGDMRAANLWPSPLMIKRSKKNSLALSLTADQMVSALLDAEPPIL
                     YSEYDPTRPFSEASMMGLLTNLADRELVHMINWAKRVPGFVDLTLHDQVHLLECAWLEI
                     LMIGLVWRSMEHPVKLLFAPNLLLDRNQGKCVEGMVEIFDMLLATSSRFRMMNLQGEEF
                     VCLKSIILLNSGVYTFLSSTLKSLEEKDHIHRVLDKITDTLIHLMAKAGLTLQQQHQRL
                     AQLLLILSHIRHMSNKGMEHLYSMKCKNVVPLYDLLLEAADAHRLHAPTSRGGASVEET
                     DQSHLATAGSTSSHSLQKYYITGEAEGFPAT"
     primer_bind     complement(1539..1574)
                     /label=A1051ERT2 BsaI mut in fusion -r
                     /note="color: black; sequence: 
                     gccagacgagGccaatcatcaggatctctagccagg; added: 2022-08-11"
     primer_bind     1555..1584
                     /label=A1050ERT2 BsaI mut in fusion -f
                     /note="color: black; sequence: 
                     gatgattggCctcgtctggcgctccatgga; added: 2022-08-11"
     primer_bind     2162..2207
                     /label=A1149 BsaI-CreER-vec-f
                     /note="color: black; sequence: 
                     ggtttccctgccacagcttaatctagaactatagtgagtcgtatta; added: 
                     2022-08-09"
     primer_bind     complement(2162..2207)
                     /label=A1152 CreER-BsaI-ins-r
                     /note="color: black; sequence: 
                     taatacgactcactatagttctagattaagctgtggcagggaaacc; added: 
                     2022-08-09"
     primer_bind     complement(2162..2182)
                     /label=ERT2 SacI -r
                     /note="color: black; sequence: 
                     cgagagctcttaagctgtggcagggaaacc"
     promoter        complement(2189..2207)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /note="color: #ffffff; direction: LEFT"
     polyA_signal    2212..2346
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
                     /note="color: #a6acb3"
     primer_bind     complement(2390..2423)
                     /label=SV40-pA-BsaI-CTCA-r
                     /note="color: black; sequence: 
                     gatggtctctTGAGgaattaaaaaacctcccacacct; added: 2022-08-02"
     CDS             2783..3574
                     /codon_start=1
                     /gene="aadA"
                     /product="aminoglycoside adenylyltransferase (Murphy, 
                     1985)"
                     /label=SmR
                     /note="confers resistance to spectinomycin and 
                     streptomycin"
                     /note="color: #ccffcc"
                     /translation="MREAVIAEVSTQLSEVVGVIERHLEPTLLAVHLYGSAVDGGLKPH
                     SDIDLLVTVTVRLDETTRRALINDLLETSASPGESEILRAVEVTIVVHDDIIPWRYPAK
                     RELQFGEWQRNDILAGIFEPATIDIDLAILLTKAREHSVALVGPAAEELFDPVPEQDLF
                     EALNETLTLWNSPPDWAGDERNVVLTLSRIWYSAVTGKIAPKDVAADWAMERLPAQYQP
                     VILEARQAYLGQEEDRLASRADQLEEFVHYVKGEITKVVGK"
     rep_origin      3667..4255
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
                     /note="color: #ffff00"
     primer_bind     4156..4175
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
                     /note="color: black; sequence: GGGAAACGCCTGGTATCTTT"
     primer_bind     4409..4426
                     /label=A757 L4440
                     /note="L4440 vector, forward primer"
                     /note="color: black; sequence: AGCGAGTCAGTGAGCGAG"
     terminator      4585..4612
                     /label=rrnB T2 terminator
                     /note="transcription terminator T2 from the E. coli rrnB 
                     gene"
                     /note="color: #ffffff"
ORIGIN
        1 atttgtccta ctcaggagag cgttcaccga caaacaacag ataaaacgaa aggcccagtc
       61 ttccgactga gcctttcgtt ttatttgatg cctggcagtt ccctactctc gcgttaacgc
      121 tagcatggat gttttcccag tcacgacgtt gtaaaacgac ggccagtctt aagctcgggc
      181 ccggtctcta gtcgccacca tgtccaattt actgaccgta caccaaaatt tgcctgcatt
      241 accggtcgat gcaacgagtg atgaggttcg caagaacctg atggacatgt tcagggatcg
      301 ccaggcgttt tctgagcata cctggaaaat gcttctgtcc gtttgccggt cgtgggcggc
      361 atggtgcaag ttgaataacc ggaaatggtt tcccgcagaa cctgaagatg ttcgcgatta
      421 tcttctatat cttcaggcgc gcggtctggc agtaaaaact atccagcaac atttgggcca
      481 gctaaacatg cttcatcgtc ggtccgggct gccacgacca agtgacagca atgctgtttc
      541 actggttatg cggcggatcc gaaaagaaaa cgttgatgcc ggtgaacgtg caaaacaggc
      601 tctagcgttc gaacgcactg atttcgacca ggttcgttca ctcatggaaa atagcgatcg
      661 ctgccaggat atacgtaatc tggcatttct ggggattgct tataacaccc tgttacgtat
      721 agccgaaatt gccaggatca gggttaaaga tatctcacgt actgacggtg ggagaatgtt
      781 aatccatatt ggcagaacga aaacgctggt tagcaccgca ggtgtagaga aggcacttag
      841 cctgggggta actaaactgg tcgagcgatg gatttccgtc tctggtgtag ctgatgatcc
      901 gaataactac ctgttttgcc gggtcagaaa aaatggtgtt gccgcgccat ctgccaccag
      961 ccagctatca actcgcgccc tggaagggat ttttgaagca actcatcgat tgatttacgg
     1021 cgctaaggat gactctggtc agagatacct ggcctggtct ggacacagtg cccgtgtcgg
     1081 agccgcgcga gatatggccc gcgctggagt ttcaataccg gagatcatgc aagctggtgg
     1141 ctggaccaat gtaaatattg tcatgaacta tatccgtaac ctggatagtg aaacaggggc
     1201 aatggtgcgc ctgctggaag atggcgatct cgagccatct gctggagaca tgagagctgc
     1261 caacctttgg ccaagcccgc tcatgatcaa acgctctaag aagaacagcc tggccttgtc
     1321 cctgacggcc gaccagatgg tcagtgcctt gttggatgct gagcccccca tactctattc
     1381 cgagtatgat cctaccagac ccttcagtga agcttcgatg atgggcttac tgaccaacct
     1441 ggcagacagg gagctggttc acatgatcaa ctgggcgaag agggtgccag gctttgtgga
     1501 tttgaccctc catgatcagg tccaccttct agaatgtgcc tggctagaga tcctgatgat
     1561 tggcctcgtc tggcgctcca tggagcaccc agtgaagcta ctgtttgctc ctaacttgct
     1621 cttggacagg aaccagggaa aatgtgtaga gggcatggtg gagatcttcg acatgctgct
     1681 ggctacatca tctcggttcc gcatgatgaa tctgcaggga gaggagtttg tgtgcctcaa
     1741 atctattatt ttgcttaatt ctggagtgta cacatttctg tccagcaccc tgaagtctct
     1801 ggaagagaag gaccatatcc accgagtcct ggacaagatc acagacactt tgatccacct
     1861 gatggccaag gcaggcctga ccctgcagca gcagcaccag cggctggccc agctcctcct
     1921 catcctctcc cacatcaggc acatgagtaa caaaggcatg gagcatctgt acagcatgaa
     1981 gtgcaagaac gtggtgcccc tctatgacct gctgctggag gcggcggacg cccaccgcct
     2041 acatgcgccc actagccgtg gaggggcatc cgtggaggag acggaccaaa gccacttggc
     2101 cactgcgggc tctacttcat cgcattcctt gcaaaagtat tacatcacgg gggaggcaga
     2161 gggtttccct gccacagctt aatctagaac tatagtgagt cgtattacgt agatccagac
     2221 atgataagat acattgatga gtttggacaa accacaacta gaatgcagtg aaaaaaatgc
     2281 tttatttgtg aaatttgtga tgctattgct ttatttgtaa ccattataag ctgcaataaa
     2341 caagttaaca acaacaattg cattcatttt atgtttcagg ttcaggggga ggtgtgggag
     2401 gttttttaat tcctcaagag acctctagac cagccaggac agaaatgcct cgacttcgct
     2461 gctacccaag gttgccgggt gacgcacacc gtggaaacgg atgaaggcac gaacccagtg
     2521 gacataagcc tgttcggttc gtaagctgta atgcaagtag cgtatgcgct cacgcaactg
     2581 gtccagaacc ttgaccgaac gcagcggtgg taacggcgca gtggcggttt tcatggcttg
     2641 ttatgactgt ttttttgggg tacagtctat gcctcgggca tccaagcagc aagcgcgtta
     2701 cgccgtgggt cgatgtttga tgttatggag cagcaacgat gttacgcagc agggcagtcg
     2761 ccctaaaaca aagttaaaca ttatgaggga agcggtgatc gccgaagtat cgactcaact
     2821 atcagaggta gttggcgtca tcgagcgcca tctcgaaccg acgttgctgg ccgtacattt
     2881 gtacggctcc gcagtggatg gcggcctgaa gccacacagt gatattgatt tgctggttac
     2941 ggtgaccgta aggcttgatg aaacaacgcg gcgagctttg atcaacgacc ttttggaaac
     3001 ttcggcttcc cctggagaga gcgagattct ccgcgctgta gaagtcacca ttgttgtgca
     3061 cgacgacatc attccgtggc gttatccagc taagcgcgaa ctgcaatttg gagaatggca
     3121 gcgcaatgac attcttgcag gtatcttcga gccagccacg atcgacattg atctggctat
     3181 cttgctgaca aaagcaagag aacatagcgt tgccttggta ggtccagcgg cggaggaact
     3241 ctttgatccg gttcctgaac aggatctatt tgaggcgcta aatgaaacct taacgctatg
     3301 gaactcgccg cccgactggg ctggcgatga gcgaaatgta gtgcttacgt tgtcccgcat
     3361 ttggtacagc gcagtaaccg gcaaaatcgc gccgaaggat gtcgctgccg actgggcaat
     3421 ggagcgcctg ccggcccagt atcagcccgt catacttgaa gctagacagg cttatcttgg
     3481 acaagaagaa gatcgcttgg cctcgcgcgc agatcagttg gaagaatttg tccactacgt
     3541 gaaaggcgag atcaccaagg tagtcggcaa ataaccctcg agccacccat gaccaaaatc
     3601 ccttaacgtg agttacgcgt cgttccactg agcgtcagac cccgtagaaa agatcaaagg
     3661 atcttcttga gatccttttt ttctgcgcgt aatctgctgc ttgcaaacaa aaaaaccacc
     3721 gctaccagcg gtggtttgtt tgccggatca agagctacca actctttttc cgaaggtaac
     3781 tggcttcagc agagcgcaga taccaaatac tgtccttcta gtgtagccgt agttaggcca
     3841 ccacttcaag aactctgtag caccgcctac atacctcgct ctgctaatcc tgttaccagt
     3901 ggctgctgcc agtggcgata agtcgtgtct taccgggttg gactcaagac gatagttacc
     3961 ggataaggcg cagcggtcgg gctgaacggg gggttcgtgc acacagccca gcttggagcg
     4021 aacgacctac accgaactga gatacctaca gcgtgagcat tgagaaagcg ccacgcttcc
     4081 cgaagggaga aaggcggaca ggtatccggt aagcggcagg gtcggaacag gagagcgcac
     4141 gagggagctt ccagggggaa acgcctggta tctttatagt cctgtcgggt ttcgccacct
     4201 ctgacttgag cgtcgatttt tgtgatgctc gtcagggggg cggagcctat ggaaaaacgc
     4261 cagcaacgcg gcctttttac ggttcctggc cttttgctgg ccttttgctc acatgttctt
     4321 tcctgcgtta tcccctgatt ctgtggataa ccgtattacc gcctttgagt gagctgatac
     4381 cgctcgccgc agccgaacga ccgagcgcag cgagtcagtg agcgaggaag cggaagagcg
     4441 cccaatacgc aaaccgcctc tccccgcgcg ttggccgatt cattaatgca gctggcacga
     4501 caggtttccc gactggaaag cgggcagtga gcgcaacgca attaatacgc gtaccgctag
     4561 ccaggaagag tttgtagaaa cgcaaaaagg ccatccgtca ggatggcctt ctgcttagtt
     4621 tgatgcctgg cagtttatgg cgggcgtcct gcccgccacc ctccgggccg ttgcttcaca
     4681 acgttcaaat ccgctcccgg cgg
//
