LOCUS       40924_170       10080 bp DNA     circular SYN 13-JAN-2022
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 10080)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 10080)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..10080
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     misc_feature    33..836
                     /label=HA-L
                     /note="left homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     misc_feature    843..868
                     /label=SA
                     /note="splice acceptor site"
     CDS             892..945
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
     CDS             961..1761
                     /label=NeoR/KanR
                     /note="aminoglycoside phosphotransferase from Tn5"
     polyA_signal    1836..2291
                     /label=PGK poly(A) signal
                     /note="mouse phosphoglycerate kinase 1 polyadenylation
                     signal"
     enhancer        2389..2692
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2694..2971
                     /note="chicken beta-actin promoter"
     intron          2973..3990
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             4090..4833
                     /label=rtTA-Advanced
                     /note="improved tetracycline-controlled transactivator"
     polyA_signal    complement(4846..4980)
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5310..6146
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     promoter        complement(6187..6205)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6212..6228)
                     /label=M13 fwd
                     /note="M13 fwd"
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6366..6665
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(8068..8925)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      9049..9637
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9925..9946
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9961..9991
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9999..10015
                     /label=lac repressor encoded by lacI binding site
                     /bound_moiety="lac repressor encoded by lacI"
                     /note="lac operator"
                     /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     10023..10039
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        10060..10078
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
ORIGIN
        1 ctagtcctgc aggtttaaac gaattcgccc tttgctttct ctgaccagca ttctctcccc
       61 tgggcctgtg ccgctttctg tctgcagctt gtggcctggg tcacctctac ggctggccca
      121 gatccttccc tgccgcctcc ttcaggttcc gtcttcctcc actccctctt ccccttgctc
      181 tctgctgtgt tgctgcccaa ggatgctctt tccggagcac ttccttctcg gcgctgcacc
      241 acgtgatgtc ctctgagcgg atcctccccg tgtctgggtc ctctccgggc atctctcctc
      301 cctcacccaa ccccatgccg tcttcactcg ctgggttccc ttttccttct ccttctgggg
      361 cctgtgccat ctctcgtttc ttaggatggc cttctccgac ggatgtctcc cttgcgtccc
      421 gcctcccctt cttgtaggcc tgcatcatca ccgtttttct ggacaacccc aaagtacccc
      481 gtctccctgg ctttagccac ctctccatcc tcttgctttc tttgcctgga caccccgttc
      541 tcctgtggat tcgggtcacc tctcactcct ttcatttggg cagctcccct acccccctta
      601 cctctctagt ctgtgctagc tcttccagcc ccctgtcatg gcatcttcca ggggtccgag
      661 agctcagcta gtcttcttcc tccaacccgg gcccctatgt ccacttcagg acagcatgtt
      721 tgctgcctcc agggatcctg tgtccccgag ctgggaccac cttatattcc cagggccggt
      781 taatgtggct ctggttctgg gtacttttat ctgtcccctc caccccacag tggggcaagc
      841 ttctgacctc ttctcttcct cccacagggc ctcgagagat ctggcagcgg agagggcaga
      901 ggaagtcttc taacatgcgg tgacgtggag gagaatcccg gccctaggct cgagacgcgt
      961 atgggatcgg ccattgaaca agatggattg cacgcaggtt ctccggccgc ttgggtggag
     1021 aggctattcg gctatgactg ggcacaacag acaatcggct gctctgatgc cgccgtgttc
     1081 cggctgtcag cgcaggggcg cccggttctt tttgtcaaga ccgacctgtc cggtgccctg
     1141 aatgaactgc aggacgaggc agcgcggcta tcgtggctgg ccacgacggg cgttccttgc
     1201 gcagctgtgc tcgacgttgt cactgaagcg ggaagggact ggctgctatt gggcgaagtg
     1261 ccggggcagg atctcctgtc atctcacctt gctcctgccg agaaagtatc catcatggct
     1321 gatgcaatgc ggcggctgca tacgcttgat ccggctacct gcccattcga ccaccaagcg
     1381 aaacatcgca tcgagcgagc acgtactcgg atggaagccg gtcttgtcga tcaggatgat
     1441 ctggacgaag agcatcaggg gctcgcgcca gccgaactgt tcgccaggct caaggcgcgc
     1501 atgcccgacg gcgaggatct cgtcgtgacc catggcgatg cctgcttgcc gaatatcatg
     1561 gtggaaaatg gccgcttttc tggattcatc gactgtggcc ggctgggtgt ggcggaccgc
     1621 tatcaggaca tagcgttggc tacccgtgat attgctgaag agcttggcgg cgaatgggct
     1681 gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt cgcagcgcat cgccttctat
     1741 cgccttcttg acgagttctt ctgaggggat ccgctgtaag tctgcagaaa ttgatgatct
     1801 attaaacaat aaagatgtcc actaaaagga agtttttctg cagaaattga tgatctatta
     1861 aacaataaag atgtccacta aaatgaagtt ttttactgtc atactttgtt aagaagggtg
     1921 agaacagagt acctacattt tgaatggaag gattggagct acgggggtgg gggtggggtg
     1981 ggattagata aatgcctgct ctttactgaa ggctctttac tattgcttta tgataatgtt
     2041 tcatagttgg atatcataat ttaaacaagc aaaaccaaat taagggccag ctcattcctc
     2101 ccactcatga tctatagatc tatagatctc tcgtgggatc attgtttttc tcttgattcc
     2161 cactttgtgg ttctaagtac tgtggtttcc aaatgtgtca gtttcatagc ctgaagaacg
     2221 agatcagcag cctctgttcc acatacactt cattctcagt attgttttgc caagttctaa
     2281 ttccatcaga agctgggagc tccaccgcgg tggcggccgc tctagaacta gttattaata
     2341 gtaatcaatt acggggtcat tagttcatag cccatatatg gagttccgcg ttacataact
     2401 tacggtaaat ggcccgcctg gctgaccgcc caacgacccc cgcccattga cgtcaataat
     2461 gacgtatgtt cccatagtaa cgccaatagg gactttccat tgacgtcaat gggtggagta
     2521 tttacggtaa actgcccact tggcagtaca tcaagtgtat catatgccaa gtacgccccc
     2581 tattgacgtc aatgacggta aatggcccgc ctggcattat gcccagtaca tgaccttatg
     2641 ggactttcct acttggcagt acatctacgt attagtcatc gctattacca tggtcgaggt
     2701 gagccccacg ttctgcttca ctctccccat ctcccccccc tccccacccc caattttgta
     2761 tttatttatt ttttaattat tttgtgcagc gatgggggcg gggggggggg gggggcgcgc
     2821 gccaggcggg gcggggcggg gcgaggggcg gggcggggcg aggcggagag gtgcggcggc
     2881 agccaatcag agcggcgcgc tccgaaagtt tccttttatg gcgaggcggc ggcggcggcg
     2941 gccctataaa aagcgaagcg cgcggcgggc ggggagtcgc tgcgacgctg ccttcgcccc
     3001 gtgccccgct ccgccgccgc ctcgcgccgc ccgccccggc tctgactgac cgcgttactc
     3061 ccacaggtga gcgggcggga cggcccttct cctccgggct gtaattagcg cttggtttaa
     3121 tgacggcttg tttcttttct gtggctgcgt gaaagccttg aggggctccg ggagggccct
     3181 ttgtgcgggg ggagcggctc ggggggtgcg tgcgtgtgtg tgtgcgtggg gagcgccgcg
     3241 tgcggctccg cgctgcccgg cggctgtgag cgctgcgggc gcggcgcggg gctttgtgcg
     3301 ctccgcagtg tgcgcgaggg gagcgcggcc gggggcggtg ccccgcggtg cggggggggc
     3361 tgcgagggga acaaaggctg cgtgcggggt gtgtgcgtgg gggggtgagc agggggtgtg
     3421 ggcgcgtcgg tcgggctgca accccccctg cacccccctc cccgagttgc tgagcacggc
     3481 ccggcttcgg gtgcggggct ccgtacgggg cgtggcgcgg ggctcgccgt gccgggcggg
     3541 gggtggcggc aggtgggggt gccgggcggg gcggggccgc ctcgggccgg ggagggctcg
     3601 ggggaggggc gcggcggccc ccggagcgcc ggcggctgtc gaggcgcggc gagccgcagc
     3661 cattgccttt tatggtaatc gtgcgagagg gcgcagggac ttcctttgtc ccaaatctgt
     3721 gcggagccga aatctgggag gcgccgccgc accccctcta gcgggcgcgg ggcgaagcgg
     3781 tgcggcgccg gcaggaagga aatgggcggg gagggccttc gtgcgtcgcc gcgccgccgt
     3841 ccccttctcc ctctccagcc tcggggctgt ccgcgggggg acggctgcct tcggggggga
     3901 cggggcaggg cggggttcgg cttctggcgt gtgaccggcg gctctagagc ctctgctaac
     3961 catgttcatg ccttcttctt tttcctacag ctcctgggca acgtgctggt tattgtgctg
     4021 tctcatcatt ttggcaaaga attcggcttg atcgaagcta gtggatcccc cgggctgcag
     4081 gaattcacca tgtctagact ggacaagagc aaagtcataa acggcgctct ggaattactc
     4141 aatggagtcg gtatcgaagg cctgacgaca aggaaactcg ctcaaaagct gggagttgag
     4201 cagcctaccc tgtactggca cgtgaagaac aagcgggccc tgctcgatgc cctgccaatc
     4261 gagatgctgg acaggcatca tacccacttc tgccccctgg aaggcgagtc atggcaagac
     4321 tttctgcgga acaacgccaa gtcattccgc tgtgctctcc tctcacatcg cgacggggct
     4381 aaagtgcatc tcggcacccg cccaacagag aaacagtacg aaaccctgga aaatcagctc
     4441 gcgttcctgt gtcagcaagg cttctccctg gagaacgcac tgtacgctct gtccgccgtg
     4501 ggccacttta cactgggctg cgtattggag gaacaggagc atcaagtagc aaaagaggaa
     4561 agagagacac ctaccaccga ttctatgccc ccacttctga gacaagcaat tgagctgttc
     4621 gaccggcagg gagccgaacc tgccttcctt ttcggcctgg aactaatcat atgtggcctg
     4681 gagaaacagc taaagtgcga aagcggcggg ccggccgacg cccttgacga ttttgactta
     4741 gacatgctcc cagccgatgc ccttgacgac tttgaccttg atatgctgcc tgctgacgct
     4801 cttgacgatt ttgaccttga catgctcccc gggtaactaa gtaaggatcc agacatgata
     4861 agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa atgctttatt
     4921 tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa taaacaagtt
     4981 aacaacaaca attgcattca ttttatgttt caggttcagg gggaggtgtg ggaggttttt
     5041 taaagcaagt aaaacctcta caaatgtggt atggctgatt atgatcctgc aagcctcgtc
     5101 gtctggccgg accacgctat ctgtgcaagg tccccggacg cgcgctccat gagcagagcg
     5161 cccgccgccg aggcaagact cgggcggcgc cctgcccgtc ccaccaggtc aacaggcggt
     5221 aaccggcctc ttcatcggga atgcgcgcga ccttcagcat cgccggcatg tcccctggcg
     5281 gacgggaagt atcagctcga ccaagcttta ctagggacag gattggtgac agaaaagccc
     5341 catccttagg cctcctcctt cctagtctcc tgatattggg tctaaccccc acctcctgtt
     5401 aggcagattc cttatctggt gacacacccc catttcctgg agccatctct ctccttgcca
     5461 gaacctctaa ggtttgctta cgatggagcc agagaggatc ctgggaggga gagcttggca
     5521 gggggtggga gggaaggggg ggatgcgtga cctgcccggt tctcagtggc caccctgcgc
     5581 taccctctcc cagaacctga gctgctctga cgcggctgtc tggtgcgttt cactgatcct
     5641 ggtgctgcag cttccttaca cttcccaaga ggagaagcag tttggaaaaa caaaatcaga
     5701 ataagttggt cctgagttct aactttggct cttcaccttt ctagtcccca atttatattg
     5761 ttcctccgtg cgtcagtttt acctgtgaga taaggccagt agccagcccc gtcctggcag
     5821 ggctgtggtg aggagggggg tgtccgtgtg gaaaactccc tttgtgagaa tggtgcgtcc
     5881 taggtgttca ccaggtcgtg gccgcctcta ctccctttct ctttctccat ccttctttcc
     5941 ttaaagagtc cccagtgcta tctgggacat attcctccgc ccagagcagg gtcccgcttc
     6001 cctaaggccc tgctctgggc ttctgggttt gagtccttgg caagcccagg agaggcgctc
     6061 aggcttccct gtcccccttc ctcgtccacc atctcatgcc cctggctctc ctgccccttc
     6121 cctacagggg ttcctggctc tgctctaagg gcaagggcga attcgcggcc gctaaattca
     6181 attcgcccta tagtgagtcg tattacaatt cactggccgt cgttttacaa cgtcgtgact
     6241 gggaaaaccc tggcgttacc caacttaatc gccttgcagc acatccccct ttcgccagct
     6301 ggcgtaatag cgaagaggcc cgcaccgatc gcccttccca acagttgcgc agcctatacg
     6361 tacggcagtt taaggtttac acctataaaa gagagagccg ttatcgtctg tttgtggatg
     6421 tacagagtga tattattgac acgccggggc gacggatggt gatccccctg gccagtgcac
     6481 gtctgctgtc agataaagtc tcccgtgaac tttacccggt ggtgcatatc ggggatgaaa
     6541 gctggcgcat gatgaccacc gatatggcca gtgtgccggt ctccgttatc ggggaagaag
     6601 tggctgatct cagccaccgc gaaaatgaca tcaaaaacgc cattaacctg atgttctggg
     6661 gaatataaat gtcaggcatg agattatcaa aaaggatctt cacctagatc cttttcacgt
     6721 agaaagccag tccgcagaaa cggtgctgac cccggatgaa tgtcagctac tgggctatct
     6781 ggacaaggga aaacgcaagc gcaaagagaa agcaggtagc ttgcagtggg cttacatggc
     6841 gatagctaga ctgggcggtt ttatggacag caagcgaacc ggaattgcca gctggggcgc
     6901 cctctggtaa ggttgggaag ccctgcaaag taaactggat ggctttcttg ccgccaagga
     6961 tctgatggcg caggggatca agctctgatc aagagacagg atgaggatcg tttcgcatga
     7021 ttgaacaaga tggattgcac gcaggttctc cggccgcttg ggtggagagg ctattcggct
     7081 atgactgggc acaacagaca atcggctgct ctgatgccgc cgtgttccgg ctgtcagcgc
     7141 aggggcgccc ggttcttttt gtcaagaccg acctgtccgg tgccctgaat gaactgcaag
     7201 acgaggcagc gcggctatcg tggctggcca cgacgggcgt tccttgcgca gctgtgctcg
     7261 acgttgtcac tgaagcggga agggactggc tgctattggg cgaagtgccg gggcaggatc
     7321 tcctgtcatc tcaccttgct cctgccgaga aagtatccat catggctgat gcaatgcggc
     7381 ggctgcatac gcttgatccg gctacctgcc cattcgacca ccaagcgaaa catcgcatcg
     7441 agcgagcacg tactcggatg gaagccggtc ttgtcgatca ggatgatctg gacgaagagc
     7501 atcaggggct cgcgccagcc gaactgttcg ccaggctcaa ggcgagcatg cccgacggcg
     7561 aggatctcgt cgtgacccat gcatggcgat gcctgcttgc cgaatatcat ggtggaaaat
     7621 ggccgctttt ctggattcat cgactgtggc cggctgggtg tggcggaccg ctatcaggac
     7681 atagcgttgg ctacccgtga tattgctgaa gagcttggcg gcgaatgggc tgaccgcttc
     7741 ctcgtgcttt acggtatcgc cgctcccgat tcgcagcgca tcgccttcta tcgccttctt
     7801 gacgagttct tctgaattat taacgcttac aatttcctga tgcggtattt tctccttacg
     7861 catctgtgcg gtatttcaca ccgcatcagg tggcactttt cggggaaatg tgcgcggaac
     7921 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gattatcaaa
     7981 aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat
     8041 atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc
     8101 gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat
     8161 acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc
     8221 ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc
     8281 tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag
     8341 ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg
     8401 ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg
     8461 atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag
     8521 taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt
     8581 catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga
     8641 atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc
     8701 acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc
     8761 aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc
     8821 ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc
     8881 cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca
     8941 atattattga agcatttatc agggttattg tctcatgacc aaaatccctt aacgtgagtt
     9001 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     9061 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     9121 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     9181 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     9241 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     9301 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     9361 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     9421 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     9481 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     9541 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     9601 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     9661 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     9721 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     9781 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     9841 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     9901 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     9961 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
    10021 cacaggaaac agctatgacc atgattacgc caagctcaga attaaccctc actaaaggga
//
