LOCUS       Exported                7151 bp ds-DNA     circular SYN 15-OCT-2020
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  recombinant plasmid
REFERENCE   1  (bases 1 to 7151)
  AUTHORS   Martin Myers
  TITLE     Direct Submission
  JOURNAL   Exported Thursday, Jan 28, 2021 from SnapGene Viewer 5.2.4
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7151
                     /organism="recombinant plasmid"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   12..141
                     /label=AAV2 ITR
     primer_bind     62..72
                     /label=ps211
                     /note="vwa5a 5pF1,use w/R2 (214) 493bp"
     protein_bind    298..568
                     /label=tetracycline response element
                     /bound_moiety="tetracycline repressor TetR"
                     /note="contains seven copies of the tetracycline operator 
                     tetO"
     promoter        601..638
                     /label=minimal CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     612..632
                     /label=ps283
                     /note="cmv minimal"
     primer_bind     763..774
                     /label=ps277
                     /note="WTHN TVA-mcherry-oG F SAlI"
     protein_bind    773..806
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    817..850
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     primer_bind     912..936
                     /label=Primer 1
     primer_bind     1424..1433
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     CDS             2708..3412
                     /codon_start=1
                     /product="monomeric derivative of DsRed fluorescent protein
                     (Shaner et al., 2004)"
                     /label=mCherry
                     /note="mammalian codon-optimized"
                     /translation="MVSKGEEDNMAIIKEFMRFKVHMEGSVNGHEFEIEGEGEGRPYEG
                     TQTAKLKVTKGGPLPFAWDILSPQFMYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNF
                     EDGGVVTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASSERMYPEDGALK
                     GEIKQRLKLKDGGHYDAEVKTTYKAKKPVQLPGAYNVNIKLDITSHNEDYTIVEQYERA
                     EGRHSTGGMDELY"
     primer_bind     complement(3395..3418)
                     /label=ps271
                     /note="WTHN hM3Dq F bsrg1"
     primer_bind     3395..3414
                     /label=PS310
                     /note="3p mCherry out"
     primer_bind     complement(3399..3424)
                     /label=Primer 2
     protein_bind    complement(3454..3487)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     protein_bind    complement(3498..3531)
                     /label=lox2272
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GGATACTT). lox2272 sites are compatible with each
                     other, but incompatible with loxP or loxN sites."
     misc_feature    3555..4143
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(3608..3628)
                     /label=WPRE-R
                     /note="WPRE, reverse primer"
     misc_feature    complement(3612..3631)
                     /label=Feature 21
     primer_bind     complement(3612..3631)
                     /label=ps125
                     /note="Ai65 PCS seq R1"
     primer_bind     complement(3630..3649)
                     /label=ps145
                     /note="Rosa26TdT (007909) Mutant Reverse (WPRE)"
     primer_bind     complement(3648..3671)
                     /label=ps126
                     /note="Ai65 PCS seq R2"
     primer_bind     complement(3722..3741)
                     /label=ps178
                     /note="P18 R CRE-dsGFP deepseq"
     primer_bind     complement(3882..3897)
                     /label=ps124
                     /note="R Ai65 MluI test"
     CDS             complement(4026..4037)
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(4178..4195)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    4184..4391
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     complement(4200..4219)
                     /label=ps176
                     /note="P1 R CRE-dsGFP deepseq"
     repeat_region   4414..4554
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     repeat_region   4414..4543
                     /label=AAV2 ITR
     primer_bind     complement(4483..4493)
                     /label=ps211
                     /note="vwa5a 5pF1,use w/R2 (214) 493bp"
     rep_origin      4629..5084
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(4716..4735)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4926..4947
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(5101..5120)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     5220..5242
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(5280..5298)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        5366..5470
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5471..6331
                     /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"
     primer_bind     complement(5689..5708)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      6502..7090
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6991..7010
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgaaacaa ttctgcagga attcggggcc
      181 gcggaggctg gatcggtccc ggtgtcttct atggaggtca aaacagcgtg gatggcgtct
      241 ccaggcgatc tgacggttca ctaaacgagc tctgcttata taggtcgagt ttaccactcc
      301 ctatcagtga tagagaaaag tgaaagtcga gtttaccact ccctatcagt gatagagaaa
      361 agtgaaagtc gagtttacca ctccctatca gtgatagaga aaagtgaaag tcgagtttac
      421 cactccctat cagtgataga gaaaagtgaa agtcgagttt accactccct atcagtgata
      481 gagaaaagtg aaagtcgagt ttaccactcc ctatcagtga tagagaaaag tgaaagtcga
      541 gtttaccact ccctatcagt gatagagaaa agtgaaagtc gagctcggta cccgggtcga
      601 gtaggcgtgt acggtgggag gcctatataa gcagagctcg tttagtgaac cgtcagatcg
      661 cctggagacg ccatccacgc tgttttgacc tccatagaag acaccgggac cgatccagcc
      721 tccgcggccc cgaattcgag ctcggtaccc ggggatcctc tagtcgacgt cgataacttc
      781 gtatagcata cattatacga agttatccga atcgcaataa cttcgtatag gatactttat
      841 acgaagttat ttggcgcgcc aaatttaaat aacgcaacgt accttcgagg gggcccaagg
      901 aattcttact tgtacgatga ccttgcacaa taacagtaca acctcgcctt tgtttccaaa
      961 catcagctcc tcctggatac acagcccctc cgatgcaggg ctgcccccgg gaaccgtcac
     1021 tcatttcggc agctacaatg tttctcgagc agctggcaat ttctcctctc cagacggtac
     1081 caccgatgac cctctgggag gtcataccgt ctggcaagtg gtcttcatcg ctttcttaac
     1141 gggcatcctg gccttggtga ccatcatcgg caacatcctg gtaattgtgt catttaaggt
     1201 caacaagcag ctgaagacgg tcaacaacta cttcctctta agcctggcct gtgccgatct
     1261 gattatcggg gtcatttcaa tgaatctgtt tacgacctac atcatcatga atcgatgggc
     1321 cttagggaac ttggcctgtg acctctggct tgccattgac tgcgtagcca gcaatgcctc
     1381 tgttatgaat cttctggtca tcagctttga cagatacttt tccatcacga ggccgctcac
     1441 gtaccgagcc aaacgaacaa caaagagagc cggtgtgatg atcggtctgg cttgggtcat
     1501 ctcctttgtc ctttgggctc ctgccatctt gttctggcaa tactttgttg gaaagagaac
     1561 tgtgcctccg ggagagtgct tcattcagtt cctcagtgag cccaccatta cttttggcac
     1621 agccatcgct ggtttttata tgcctgtcac cattatgact attttatact ggaggatcta
     1681 taaggaaact gaaaagcgta ccaaagagct tgctggcctg caagcctctg ggacagaggc
     1741 agagacagaa aactttgtcc accccacggg cagttctcga agctgcagca gttacgaact
     1801 tcaacagcaa agcatgaaac gctccaacag gaggaagtat ggccgctgcc acttctggtt
     1861 cacaaccaag agctggaaac ccagctccga gcagatggac caagaccaca gcagcagtga
     1921 cagttggaac aacaatgatg ctgctgcctc cctggagaac tccgcctcct ccgacgagga
     1981 ggacattggc tccgagacga gagccatcta ctccatcgtg ctcaagcttc cgggtcacag
     2041 caccatcctc aactccacca agttaccctc atcggacaac ctgcaggtgc ctgaggagga
     2101 gctggggatg gtggacttgg agaggaaagc cgacaagctg caggcccaga agagcgtgga
     2161 cgatggaggc agttttccaa aaagcttctc caagcttccc atccagctag agtcagccgt
     2221 ggacacagct aagacttctg acgtcaactc ctcagtgggt aagagcacgg ccactctacc
     2281 tctgtccttc aaggaagcca ctctggccaa gaggtttgct ctgaagacca gaagtcagat
     2341 cactaagcgg aaaaggatgt ccctggtcaa ggagaagaaa gcggcccaga ccctcagtgc
     2401 gatcttgctt gccttcatca tcacttggac cccatacaac atcatggttc tggtgaacac
     2461 cttttgtgac agctgcatac ccaaaacctt ttggaatctg ggctactggc tgtgctacat
     2521 caacagcacc gtgaaccccg tgtgctatgc tctgtgcaac aaaacattca gaaccacttt
     2581 caagatgctg ctgctgtgcc agtgtgacaa aaaaaagagg cgcaagcagc agtaccagca
     2641 gagacagtcg gtcatttttc acaagcgcgc acccgagcag gccttgaagg atcccccggt
     2701 cgccaccatg gtgagcaagg gcgaggagga taacatggcc atcatcaagg agttcatgcg
     2761 cttcaaggtg cacatggagg gctccgtgaa cggccacgag ttcgagatcg agggcgaggg
     2821 cgagggccgc ccctacgagg gcacccagac cgccaagctg aaggtgacca agggtggccc
     2881 cctgcccttc gcctgggaca tcctgtcccc tcagttcatg tacggctcca aggcctacgt
     2941 gaagcacccc gccgacatcc ccgactactt gaagctgtcc ttccccgagg gcttcaagtg
     3001 ggagcgcgtg atgaacttcg aggacggcgg cgtggtgacc gtgacccagg actcctccct
     3061 gcaggacggc gagttcatct acaaggtgaa gctgcgcggc accaacttcc cctccgacgg
     3121 ccccgtaatg cagaagaaga ccatgggctg ggaggcctcc tccgagcgga tgtaccccga
     3181 ggacggcgcc ctgaagggcg agatcaagca gaggctgaag ctgaaggacg gcggccacta
     3241 cgacgctgag gtcaagacca cctacaaggc caagaagccc gtgcagctgc ccggcgccta
     3301 caacgtcaac atcaagttgg acatcacctc ccacaacgag gactacacca tcgtggaaca
     3361 gtacgaacgc gccgagggcc gccactccac cggcggcatg gacgagctgt acaagtaagc
     3421 tagctagttg gcgcgccaac ctggccggcc ttgataactt cgtataatgt atgctatacg
     3481 aagttatccg aatcgcaata acttcgtata aagtatccta tacgaagtta tcgaagatca
     3541 tcaagcttat cgataatcaa cctctggatt acaaaatttg tgaaagattg actggtattc
     3601 ttaactatgt tgctcctttt acgctatgtg gatacgctgc tttaatgcct ttgtatcatg
     3661 ctattgcttc ccgtatggct ttcattttct cctccttgta taaatcctgg ttgctgtctc
     3721 tttatgagga gttgtggccc gttgtcaggc aacgtggcgt ggtgtgcact gtgtttgctg
     3781 acgcaacccc cactggttgg ggcattgcca ccacctgtca gctcctttcc gggactttcg
     3841 ctttccccct ccctattgcc acggcggaac tcatcgccgc ctgccttgcc cgctgctgga
     3901 caggggctcg gctgttgggc actgacaatt ccgtggtgtt gtcggggaaa tcatcgtcct
     3961 ttccttggct gctcgcctat gttgccacct ggattctgcg cgggacgtcc ttctgctacg
     4021 tcccttcggc cctcaatcca gcggaccttc cttcccgcgg cctgctgccg gctctgcggc
     4081 ctcttccgcg tcttcgcctt cgccctcaga cgagtcggat ctccctttgg gccgcctccc
     4141 cgcatcgata ccgagcgctg ctcgagagat cgatctgcct cgactgtgcc ttctagttgc
     4201 cagccatctg ttgtttgccc ctcccccgtg ccttccttga ccctggaagg tgccactccc
     4261 actgtccttt cctaataaaa tgaggaaatt gcatcgcatt gtctgagtag gtgtcattct
     4321 attctggggg gtggggtggg gcaggacagc aagggggagg attgggaaga caatagcagg
     4381 catgctgggg acacgtgcgg accgagcggc cgcaggaacc cctagtgatg gagttggcca
     4441 ctccctctct gcgcgctcgc tcgctcactg aggccgggcg accaaaggtc gcccgacgcc
     4501 cgggctttgc ccgggcggcc tcagtgagcg agcgagcgcg cagctgcctg caggggcgcc
     4561 tgatgcggta ttttctcctt acgcatctgt gcggtatttc acaccgcata cgtcaaagca
     4621 accatagtac gcgccctgta gcggcgcatt aagcgcggcg ggtgtggtgg ttacgcgcag
     4681 cgtgaccgct acacttgcca gcgccctagc gcccgctcct ttcgctttct tcccttcctt
     4741 tctcgccacg ttcgccggct ttccccgtca agctctaaat cgggggctcc ctttagggtt
     4801 ccgatttagt gctttacggc acctcgaccc caaaaaactt gatttgggtg atggttcacg
     4861 tagtgggcca tcgccctgat agacggtttt tcgccctttg acgttggagt ccacgttctt
     4921 taatagtgga ctcttgttcc aaactggaac aacactcaac cctatctcgg gctattcttt
     4981 tgatttataa gggattttgc cgatttcggc ctattggtta aaaaatgagc tgatttaaca
     5041 aaaatttaac gcgaatttta acaaaatatt aacgtttaca attttatggt gcactctcag
     5101 tacaatctgc tctgatgccg catagttaag ccagccccga cacccgccaa cacccgctga
     5161 cgcgccctga cgggcttgtc tgctcccggc atccgcttac agacaagctg tgaccgtctc
     5221 cgggagctgc atgtgtcaga ggttttcacc gtcatcaccg aaacgcgcga gacgaaaggg
     5281 cctcgtgata cgcctatttt tataggttaa tgtcatgata ataatggttt cttagacgtc
     5341 aggtggcact tttcggggaa atgtgcgcgg aacccctatt tgtttatttt tctaaataca
     5401 ttcaaatatg tatccgctca tgagacaata accctgataa atgcttcaat aatattgaaa
     5461 aaggaagagt atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt
     5521 ttgccttcct gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca
     5581 gttgggtgca cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag
     5641 ttttcgcccc gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc
     5701 ggtattatcc cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca
     5761 gaatgacttg gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt
     5821 aagagaatta tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct
     5881 gacaacgatc ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt
     5941 aactcgcctt gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga
     6001 caccacgatg cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact
     6061 tactctagct tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc
     6121 acttctgcgc tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga
     6181 gcgtgggtct cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt
     6241 agttatctac acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga
     6301 gataggtgcc tcactgatta agcattggta actgtcagac caagtttact catatatact
     6361 ttagattgat ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga
     6421 taatctcatg accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt
     6481 agaaaagatc aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca
     6541 aacaaaaaaa ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct
     6601 ttttccgaag gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta
     6661 gccgtagtta ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct
     6721 aatcctgtta ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc
     6781 aagacgatag ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca
     6841 gcccagcttg gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga
     6901 aagcgccacg cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg
     6961 aacaggagag cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt
     7021 cgggtttcgc cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag
     7081 cctatggaaa aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt
     7141 tgctcacatg t
//
