LOCUS       62056_1865        6266 bp DNA     circular SYN 16-MAY-2020
DEFINITION  Cloning vector p15A-PTET-CreGTG-AAV-PBAD-FlpGTG, complete sequence.
ACCESSION   MN623122
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6266)
  AUTHORS   Guo L.
  TITLE     Direct Submission
  JOURNAL   Submitted (28-OCT-2019) Jiangnan University, State Key Laboratory of
            Food Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122,
            China
REFERENCE   2  (bases 1 to 6266)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Submitted 
            (28-OCT-2019) Jiangnan University, State Key Laboratory of Food 
            Science and Technology, 1800 Lihu Avenue, Wuxi, Jiangsu 214122, 
            China"
FEATURES             Location/Qualifiers
     source          1..6266
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     terminator      complement(6..33)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     terminator      complement(49..120)
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB
                     gene"
     CDS             complement(162..1430)
                     /codon_start=1
                     /label=FLP
                     /note="site-specific recombinase"
                     /translation="VPQFDILCKTPPKVLVRQFVERFERPSGEKIALCAAELTYLCWMI
                     THNGTAIKRATFMSYNTIISNSLSFDIVNKSLQFKYKTQKATILEASLKKLIPAWEFTI
                     IPYYGQKHQSDITDIVSSLQLQFESSEEADKGNSHSKKMLKALLSEGESIWEITEKILN
                     SFEYTSRFTKTKTLYQFLFLATFINCGRFSDIKNVDPKSFKLVQNKYLGVIIQCLVTET
                     KTSVSRHIYFFSARGRIDPLVYLDEFLRNSEPVLKRVNRTGNSSSNKQEYQLLKDNLVR
                     SYNKALKKNAPYSIFAIKNGPKSHIGRHLMTSFLSMKGLTELTNVVGNWSDKRASAVAR
                     TTYTHQITAIPDHYFALVSRYYAYDPISKEMIALKDETNPIEEWQHIEQLKGSAEGSIR
                     YPAWNGIISQEVLDYLSSYINRRI"
     promoter        complement(1475..1759)
                     /label=araBAD promoter
                     /note="promoter of the L-arabinose operon of E. coli; the
                     araC regulatory gene is transcribed in the opposite 
                     direction (Guzman et al., 1995)"
     CDS             1786..2661
                     /codon_start=1
                     /label=araC
                     /note="L-arabinose regulatory protein"
                     /translation="MAEAQNDPLLPGYSFNAHLVAGLTPIEANGYLDFFIDRPLGMKGY
                     ILNLTIRGQGVVKNQGREFVCRPGDILLFPPGEIHHYGRHPEAREWYHQWVYFRPRAYW
                     HEWLNWPSIFANTGFFRPDEAHQPHFSDLFGQIINAGQGEGRYSELLAINLLEQLLLRR
                     MEAINESLHPPMDNRVREACQYISDHLADSNFDIASVAQHVCLSPSRLSHLFRQQLGIS
                     VLSWREDQRISQAKLLLSTTRMPIATVGRNVGFDDQLYFSRVFKKCTGASPSEFRAGCE
                     EKVNDVAVKLS"
     rep_origin      complement(2865..3410)
                     /direction=LEFT
                     /label=p15A ori
                     /note="Plasmids containing the medium-copy-number p15A
                     origin of replication can be propagated in E. coli cells 
                     that contain a second plasmid with the ColE1 origin."
     terminator      complement(3524..3618)
                     /label=lambda t0 terminator
                     /note="transcription terminator from phage lambda"
     CDS             complement(3642..4298)
                     /codon_start=1
                     /label=CmR
                     /note="chloramphenicol acetyltransferase"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQGGA"
     promoter        complement(4299..4401)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding
                     chloramphenicol acetyltransferase"
     CDS             complement(4434..5057)
                     /codon_start=1
                     /label=TetR
                     /note="tetracycline repressor TetR"
                     /translation="MMSRLDKSKVINSALELLNEVGIEGLTTRKLAQKLGVEQPTLYWH
                     VKNKRALLDALAIEMLDRHHTHFCPLEGESWQDFLRNNAKSFRCALLSHRDGAKVHLGT
                     RPTEKQYETLENQLAFLCQQGFSLENALYALSAVGHFTLGCVLEDQEHQVAKEERETPT
                     TDSMPPLLRQAIELFDHQGAEPAFLFGLELIICGLEKQLKCESGS"
     promoter        5073..5128
                     /label=tetR/tetA promoters
                     /note="overlapping promoters for bacterial tetR and tetA"
     CDS             5161..5178
                     /codon_start=1
                     /label=6xHis
                     /note="6xHis affinity tag"
                     /translation="HHHHHH"
     CDS             5188..6213
                     /codon_start=1
                     /label=Cre
                     /note="site-specific recombinase"
                     /translation="SNLLTVHQNLPALPVDATSDEVRKNLMDMFRDRQAFSEHTWKMLL
                     SVCRSWAAWCKLNNRKWFPAEPEDVRDYLLYLQARGLAVKTIQQHLGQLNMLHRRSGLP
                     RPSDSNAVSLVMRRIRKENVDAGERAKQALAFERTDFDQVRSLMENSDRCQDIRNLAFL
                     GIAYNTLLRIAEIARIRVKDISRTDGGRMLIHIGRTKTLVSTAGVEKALSLGVTKLVER
                     WISVSGVADDPNNYLFCRVRKNGVAAPSATSQLSTRALEGIFEATHRLIYGAKDDSGQR
                     YLAWSGHSARVGAARDMARAGVSIPEIMQAGGWTNVNIVMNYIRNLDSETGAMVRLLED
                     GD"
ORIGIN
        1 tcgaccgcag aaaggcccac ccgaaggtga gccagtgtga ctctagtaga gagcgttcac
       61 cgacaaacaa cagataaaac gaaaggccca gtctttcgac tgagcctttc gttttatttg
      121 atgcctggag atccttactc gagtgcggcc gcaagctttt atatgcgtct atttatgtag
      181 gatgaaaggt agtctagtac ctcctgtgat attatcccat tccatgcggg gtatcgtatg
      241 cttccttcag cactaccctt tagctgttct atatgctgcc actcctcaat tggattagtc
      301 tcatccttca atgctatcat ttcctttgat attggatcat atgcatagta ccgagaaact
      361 agtgcgaagt agtgatcagg tattgctgtt atctgatgag tatacgttgt cctggccacg
      421 gcagaagcac gcttatcgct ccaatttccc acaacattag tcaactccgt taggcccttc
      481 attgaaagaa atgaggtcat caaatgtctt ccaatgtgag attttgggcc attttttata
      541 gcaaagattg aataaggcgc atttttcttc aaagctttat tgtacgatct gactaagtta
      601 tcttttaata attggtattc ctgtttattg cttgaagaat tgccggtcct atttactcgt
      661 tttaggactg gttcagaatt cctcaaaaat tcatccaaat atacaagtgg atcgatccta
      721 ccccttgcgc taaagaagta tatgtgccta ctaacgcttg tctttgtctc tgtcactaaa
      781 cactggatta ttactcccag atacttattt tggactaatt taaatgattt cggatcaacg
      841 ttcttaatat cgctgaatct tccacaattg atgaaagtag ctaggaagag gaattggtat
      901 aaagtttttg tttttgtaaa tctcgaagta tactcaaacg aatttagtat tttctcagtg
      961 atctcccaga tgctttcacc ctcacttaga agtgctttaa gcattttttt actgtggcta
     1021 tttcccttat ctgcttcttc cgatgattcg aactgtaatt gcaaactact tacaatatca
     1081 gtgatatcag attgatgttt ttgtccatag taaggaataa ttgtaaattc ccaagcagga
     1141 atcaatttct ttaatgaggc ttccagaatt gttgcttttt gcgtcttgta tttaaactgg
     1201 agtgatttat tgacaatatc gaaactcagc gaattgctta tgatagtatt atagctcatg
     1261 aatgtggctc tcttgattgc tgttccgtta tgtgtaatca tccaacataa ataggttagt
     1321 tcagcagcac ataatgctat tttctcacct gaaggtcttt caaacctttc cacaaactga
     1381 cgaacaagca ccttaggtgg tgttttacat aatatatcaa attgtggcac agatctcttt
     1441 cctgtgtgac tgcagctagc ccaaaaaaac gggtatggag aaacagtaga gagttgcgat
     1501 aaaaagcgtc aggtaggatc cgctaatctt atggataaaa atgctatggc atagcaaagt
     1561 gtgacgccgt gcaaataatc aatgtggact tttctgccgt gattatagac acttttgtta
     1621 cgcgtttttg tcatggcttt ggtcccgctt tgttacagaa tgcttttaat aagcggggtt
     1681 accggtttgg ttagcgagaa gagccagtaa aagacgcagt gacggcaatg tctgatgcaa
     1741 tatggacaat tggtttcttc tctgaatggc gggagtatga aaagtatggc tgaagcgcaa
     1801 aatgatcccc tgctgccggg atactcgttt aatgcccatc tggtggcggg tttaacgccg
     1861 attgaggcca acggttatct cgattttttt atcgaccgac cgctgggaat gaaaggttat
     1921 attctcaatc tcaccattcg cggtcagggg gtggtgaaaa atcagggacg agaatttgtt
     1981 tgccgaccgg gtgatatttt gctgttcccg ccaggagaga ttcatcacta cggtcgtcat
     2041 ccggaggctc gcgaatggta tcaccagtgg gtttactttc gtccgcgcgc ctactggcat
     2101 gaatggctta actggccgtc aatatttgcc aatacggggt tctttcgccc ggatgaagcg
     2161 caccagccgc atttcagcga cctgtttggg caaatcatta acgccgggca aggggaaggg
     2221 cgctattcgg agctgctggc gataaatctg cttgagcaat tgttactgcg gcgcatggaa
     2281 gcgattaacg agtcgctcca tccaccgatg gataatcggg tacgcgaggc ttgtcagtac
     2341 atcagcgatc acctggcaga cagcaatttt gatatcgcca gcgtcgcaca gcatgtttgc
     2401 ttgtcgccgt cgcgtctgtc acatcttttc cgccagcagt tagggattag cgtcttaagc
     2461 tggcgcgagg accaacgtat cagccaggcg aagctgcttt tgagcaccac ccggatgcct
     2521 atcgccaccg tcggtcgcaa tgttggtttt gacgatcaac tctatttctc gcgggtattt
     2581 aaaaaatgca ccggggccag cccgagcgag ttccgtgccg gttgtgaaga aaaagtgaat
     2641 gatgtagccg tcaagttgtc ataattggta acgaatcaga caattgacgg cttgacggag
     2701 tagcataggc ctagggatat attccgcttc ctcgctcact gactcgctac gctcggtcgt
     2761 tcgactgcgg cgagcggaaa tggcttacga acggggcgga gatttcctgg aagatgccag
     2821 gaagatactt aacagggaag tgagagggcc gcggcaaagc cgtttttcca taggctccgc
     2881 ccccctgaca agcatcacga aatctgacgc tcaaatcagt ggtggcgaaa cccgacagga
     2941 ctataaagat accaggcgtt tccccctggc ggctccctcg tgcgctctcc tgttcctgcc
     3001 tttcggttta ccggtgtcat tccgctgtta tggccgcgtt tgtctcattc cacgcctgac
     3061 actcagttcc gggtaggcag ttcgctccaa gctggactgt atgcacgaac cccccgttca
     3121 gtccgaccgc tgcgccttat ccggtaacta tcgtcttgag tccaacccgg aaagacatgc
     3181 aaaagcacca ctggcagcag ccactggtaa ttgatttaga ggagttagtc ttgaagtcat
     3241 gcgccggtta aggctaaact gaaaggacaa gttttggtga ctgcgctcct ccaagccagt
     3301 tacctcggtt caaagagttg gtagctcaga gaaccttcga aaaaccgccc tgcaaggcgg
     3361 ttttttcgtt ttcagagcaa gagattacgc gcagaccaaa acgatctcaa gaagatcatc
     3421 ttattaatca gataaaatat ttctagattt cagtgcaatt tatctcttca aatgtagcac
     3481 ctgaagtcag ccccatacga tataagttgt tactagtgct tggattctca ccaataaaaa
     3541 acgcccggcg gcaaccgagc gttctgaaca aatccagatg gagttctgag gtcattactg
     3601 gatctatcaa caggagtcca agcgagctcg atatcaaatt acgccccgcc ctgccactca
     3661 tcgcagtact gttgtaattc attaagcatt ctgccgacat ggaagccatc acaaacggca
     3721 tgatgaacct gaatcgccag cggcatcagc accttgtcgc cttgcgtata atatttgccc
     3781 atggtgaaaa cgggggcgaa gaagttgtcc atattggcca cgtttaaatc aaaactggtg
     3841 aaactcaccc agggattggc tgagacgaaa aacatattct caataaaccc tttagggaaa
     3901 taggccaggt tttcaccgta acacgccaca tcttgcgaat atatgtgtag aaactgccgg
     3961 aaatcgtcgt ggtattcact ccagagcgat gaaaacgttt cagtttgctc atggaaaacg
     4021 gtgtaacaag ggtgaacact atcccatatc accagctcac cgtctttcat tgccatacga
     4081 aattccggat gagcattcat caggcgggca agaatgtgaa taaaggccgg ataaaacttg
     4141 tgcttatttt tctttacggt ctttaaaaag gccgtaatat ccagctgaac ggtctggtta
     4201 taggtacatt gagcaactga ctgaaatgcc tcaaaatgtt ctttacgatg ccattgggat
     4261 atatcaacgg tggtatatcc agtgattttt ttctccattt tagcttcctt agctcctgaa
     4321 aatctcgata actcaaaaaa tacgcccggt agtgatctta tttcattatg gtgaaagttg
     4381 gaacctctta cgtgccgatc aacgtctcat tttcgccaga tatcaatttc ttaagaccca
     4441 ctttcacatt taagttgttt ttctaatccg catatgatca attcaaggcc gaataagaag
     4501 gctggctctg caccttggtg atcaaataat tcgatagctt gtcgtaataa tggcggcata
     4561 ctatcagtag taggtgtttc cctttcttct ttagcgactt gatgctcttg atcttccaat
     4621 acgcaaccta aagtaaaatg ccccacagcg ctgagtgcat ataatgcatt ctctagtgaa
     4681 aaaccttgtt ggcataaaaa ggctaattga ttttcgagag tttcatactg tttttctgta
     4741 ggccgtgtac ctaaatgtac ttttgctcca tcgcgatgac ttagtaaagc acatctaaaa
     4801 cttttagcgt tattacgtaa aaaatcttgc cagctttccc cttctaaagg gcaaaagtga
     4861 gtatggtgcc tatctaacat ctcaatggct aaggcgtcga gcaaagcccg cttatttttt
     4921 acatgccaat acaatgtagg ctgctctaca cctagcttct gggcgagttt acgggttgtt
     4981 aaaccttcga ttccgacctc attaagcagc tctaatgcgc tgttaatcac tttactttta
     5041 tctaatctag acatcattaa ttcctaattt ttgttgacac tctatcgttg atagagttat
     5101 tttaccactc cctatcagtg atagagaaaa gaattcaggg acaggattaa atacgtgaaa
     5161 catcatcacc atcaccacct gcagggctcc aatttactga ccgtacacca aaatttgcct
     5221 gcattaccgg tcgatgcaac gagtgatgag gttcgcaaga acctgatgga catgttcagg
     5281 gatcgccagg cgttttctga gcatacctgg aaaatgcttc tgtccgtttg ccggtcgtgg
     5341 gcggcatggt gcaagttgaa taaccggaaa tggtttcccg cagaacctga agatgttcgc
     5401 gattatcttc tatatcttca ggcgcgcggt ctggcagtaa aaactatcca gcaacatttg
     5461 ggccagctaa acatgcttca tcgtcggtcc gggctgccac gaccaagtga cagcaatgct
     5521 gtttcactgg ttatgcggcg gatccgaaaa gaaaacgttg atgccggtga acgtgcaaaa
     5581 caggctctag cgttcgaacg cactgatttc gaccaggttc gttcactcat ggaaaatagc
     5641 gatcgctgcc aggatatacg taatctggca tttctgggga ttgcttataa caccctgtta
     5701 cgtatagccg aaattgccag gatcagggtt aaagatatct cacgtactga cggtgggaga
     5761 atgttaatcc atattggcag aacgaaaacg ctggttagca ccgcaggtgt agagaaggca
     5821 cttagcctgg gggtaactaa actggtcgag cgatggattt ccgtctctgg tgtagctgat
     5881 gatccgaata actacctgtt ttgccgggtc agaaaaaatg gtgttgccgc gccatctgcc
     5941 accagccagc tatcaactcg cgccctggaa gggatttttg aagcaactca tcgattgatt
     6001 tacggcgcta aggatgactc tggtcagaga tacctggcct ggtctggaca cagtgcccgt
     6061 gtcggagccg cgcgagatat ggcccgcgct ggagtttcaa taccggagat catgcaagct
     6121 ggtggctgga ccaatgtaaa tattgtcatg aactatatcc gtaacctgga tagtgaaaca
     6181 ggggcaatgg tgcgcctgct ggaagatggc gatgcagcaa acgacgaaaa ctacgctgct
     6241 gctgtttaat aagcttgcgg ccgcac
//
