LOCUS       Exported                5639 bp ds-DNA     circular SYN 19-SEP-2018
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5639)
  AUTHORS   Rui Lin
  TITLE     Direct Submission
  JOURNAL   Exported Sep 20, 2018 from SnapGene Viewer 4.2.4
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..5639
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     repeat_region   1..141
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     promoter        159..473
                     /label=tight TRE promoter
                     /note="Tet-responsive promoter PTight, consisting of seven 
                     tet operator sequences followed by the minimal CMV 
                     promoter"
     protein_bind    167..185
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    203..221
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    238..256
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    274..292
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    310..328
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    345..363
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    381..399
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     misc_feature    484..515
                     /label=rox1
     misc_feature    518..549
                     /label=rox3
     CDS             complement(558..1700)
                     /codon_start=1
                     /label=vCre
                     /translation="MIENQLSLLGDFSGVRPDDVKTAIQAAQKKGINVAENEQFKAAFE
                     HLLNEFKKREERYSPNTLRRLESAWTCFVDWCLANHRHSLPATPDTVEAFFIERAEELH
                     RNTLSVYRWAISRVHRVAGCPDPCLDIYVEDRLKAIARKKVREGEAVKQASPFNEQHLL
                     KLTSLWYRSDKLLLRRNLALLAVAYESMLRASELANIRVSDMELAGDGTAILTIPITKT
                     NHSGEPDTCILSQDVVSLLMDYTEAGKLDMSSDGFLFVGVSKHNTCIKPKKDKQTGEVL
                     HKPITTKTVEGVFYSAWETLDLGRQGVKPFTAHSARVGAAQDLLKKGYNTLQIQQSGRW
                     SSGAMVARYGRAILARDGAMAHSRVKTRSAPMQWGKDEKD"
     misc_feature    complement(1711..1742)
                     /label=rox1
     misc_feature    complement(1745..1776)
                     /label=rox3
     misc_feature    1789..2377
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     polyA_signal    2386..2862
                     /label=hGH poly(A) signal
                     /note="human growth hormone polyadenylation signal"
     repeat_region   2902..3042
                     /label=AAV2 ITR
                     /note="inverted terminal repeat of adeno-associated virus 
                     serotype 2"
     rep_origin      3117..3572
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        3854..3958
                     /gene="bla"
                     /label=AmpR promoter
     CDS             3959..4819
                     /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"
     rep_origin      4990..5578
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 cctgcaggca gctgcgcgct cgctcgctca ctgaggccgc ccgggcaaag cccgggcgtc
       61 gggcgacctt tggtcgcccg gcctcagtga gcgagcgagc gcgcagagag ggagtggcca
      121 actccatcac taggggttcc tgcggccgca cgcgtctcga gtttactccc tatcagtgat
      181 agagaacgta tgtcgagttt actccctatc agtgatagag aacgatgtcg agtttactcc
      241 ctatcagtga tagagaacgt atgtcgagtt tactccctat cagtgataga gaacgtatgt
      301 cgagtttact ccctatcagt gatagagaac gtatgtcgag tttatcccta tcagtgatag
      361 agaacgtatg tcgagtttac tccctatcag tgatagagaa cgtatgtcga ggtaggcgtg
      421 tacggtggga ggcctatata agcagagctc gtttagtgaa ccgtcagatc gcctggagaa
      481 ttctaacttt aaataatgcc aattatttaa agttagctaa ctttaaataa tgtcgattat
      541 ttaaagttag gcgcgcctta gtccttttcg tcctttcccc actgcattgg tgctgaccga
      601 gtcttgaccc tgctatgtgc cattgctccg tctcgagcca gaatggctcg tccatagcgt
      661 gccaccatag ccccactaga ccacctgccg gactgctgga tctgcagagt attgtatcct
      721 ttcttcagca ggtcctgtgc tgctccgact ctggcgctat gtgctgtaaa aggcttcacg
      781 ccctgccgtc ccagatccag tgtttcccag gctgaataga agaccccctc cacggttttt
      841 gtggtaatag gcttatgcag cacctcgcca gtctgtttat ctttcttggg cttgatacat
      901 gtattgtgtt tgctgactcc cacaaacagg aacccatcgg agctcatgtc cagcttgccg
      961 gcttcggtgt agtccatcag cagtgagacc acatcctgac tcaggatgca agtgtctggc
     1021 tctccagaat ggttggtctt agtaataggg attgtcagaa tagcggtccc atcgccggcc
     1081 agctccatgt cggagacgcg gatatttgcc agttcagaag ctcgcagcat ggactcataa
     1141 gccacggcca gcagggccag gtttcgcctc agcagcagtt tgtcgcttct gtaccacagt
     1201 gatgtcagct tcagcaggtg ctgttcgttg aaggggctgg cctgtttgac tgcttcgccc
     1261 tctctcactt tcttccgtgc aatagccttc agtctatcct ccacatagat atccagacaa
     1321 gggtcggggc atccagcgac cctatgcact ctggaaatgg cccagcggta cacgctcagg
     1381 gtattgcggt gcagttcctc ggccctttcg ataaagaatg cctcgacagt gtcgggtgtt
     1441 gctggcagac tatgcctgtg gttagccaga caccaatcca caaagcaggt ccaagcactc
     1501 tccagtctcc gcagagtatt tggagagtac cgttcctccc gcttcttgaa ctcgttcagc
     1561 aggtgctcaa aagcggcttt gaactgttca ttctctgcca cgttgatgcc cttcttctgt
     1621 gcggcctgaa tggcggtctt gacatcatcg ggtcggactc ctgaaaaatc tcccagcagt
     1681 gacagctggt tctcaatcat ggtggctagc taactttaaa taattggcat tatttaaagt
     1741 tagctaactt taaataatcg acattattta aagttaaagc ttatcgataa tcaacctctg
     1801 gattacaaaa tttgtgaaag attgactggt attcttaact atgttgctcc ttttacgcta
     1861 tgtggatacg ctgctttaat gcctttgtat catgctattg cttcccgtat ggctttcatt
     1921 ttctcctcct tgtataaatc ctggttgctg tctctttatg aggagttgtg gcccgttgtc
     1981 aggcaacgtg gcgtggtgtg cactgtgttt gctgacgcaa cccccactgg ttggggcatt
     2041 gccaccacct gtcagctcct ttccgggact ttcgctttcc ccctccctat tgccacggcg
     2101 gaactcatcg ccgcctgcct tgcccgctgc tggacagggg ctcggctgtt gggcactgac
     2161 aattccgtgg tgttgtcggg gaaatcatcg tcctttcctt ggctgctcgc ctgtgttgcc
     2221 acctggattc tgcgcgggac gtccttctgc tacgtccctt cggccctcaa tccagcggac
     2281 cttccttccc gcggcctgct gccggctctg cggcctcttc cgcgtcttcg ccttcgccct
     2341 cagacgagtc ggatctccct ttgggccgcc tccccgcaga tctacgggtg gcatccctgt
     2401 gacccctccc cagtgcctct cctggccctg gaagttgcca ctccagtgcc caccagcctt
     2461 gtcctaataa aattaagttg catcattttg tctgactagg tgtccttcta taatattatg
     2521 gggtggaggg gggtggtatg gagcaagggg caagttggga agacaacctg tagggcctgc
     2581 ggggtctatt gggaaccaag ctggagtgca gtggcacaat cttggctcac tgcaatctcc
     2641 gcctcctggg ttcaagcgat tctcctgcct cagcctcccg agttgttggg attccaggca
     2701 tgcatgacca ggctcagcta atttttgttt ttttggtaga gacggggttt caccatattg
     2761 gccaggctgg tctccaactc ctaatctcag gtgatctacc caccttggcc tcccaaattg
     2821 ctgggattac aggcgtgaac cactgctccc ttccctgtcc ttctgatttt gtaggtaacc
     2881 acgtgcggac cgagcggccg caggaacccc tagtgatgga gttggccact ccctctctgc
     2941 gcgctcgctc gctcactgag gccgggcgac caaaggtcgc ccgacgcccg ggctttgccc
     3001 gggcggcctc agtgagcgag cgagcgcgca gctgcctgca ggggcgcctg atgcggtatt
     3061 ttctccttac gcatctgtgc ggtatttcac accgcatacg tcaaagcaac catagtacgc
     3121 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     3181 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     3241 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
     3301 tttacggcac ctcgacccca aaaaacttga tttgggtgat ggttcacgta gtgggccatc
     3361 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     3421 cttgttccaa actggaacaa cactcaaccc tatctcgggc tattcttttg atttataagg
     3481 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3541 gaattttaac aaaatattaa cgtttacaat tttatggtgc actctcagta caatctgctc
     3601 tgatgccgca tagttaagcc agccccgaca cccgccaaca cccgctgacg cgccctgacg
     3661 ggcttgtctg ctcccggcat ccgcttacag acaagctgtg accgtctccg ggagctgcat
     3721 gtgtcagagg ttttcaccgt catcaccgaa acgcgcgaga cgaaagggcc tcgtgatacg
     3781 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     3841 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     3901 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     3961 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     4021 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     4081 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     4141 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     4201 tattgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     4261 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     4321 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     4381 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     4441 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     4501 tgtagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     4561 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     4621 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     4681 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     4741 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     4801 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     4861 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     4921 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     4981 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     5041 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     5101 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     5161 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     5221 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     5281 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     5341 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     5401 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     5461 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     5521 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     5581 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgt
//
