LOCUS       40924_130        9809 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 9809)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 9809)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..9809
                     /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             955..1551
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
     polyA_signal    1592..1816
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     enhancer        1822..2201
                     /note="CMV enhancer"
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        2203..2480
                     /note="chicken beta-actin promoter"
     intron          2482..3499
                     /label=chimeric intron
                     /note="chimera between introns from chicken beta-actin and
                     rabbit beta-globin"
     CDS             3561..4277
                     /label=EGFP
                     /note="enhanced GFP"
     polyA_signal    4435..4490
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(4851..4867)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(4875..4891)
                     /label=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)."
     promoter        complement(4899..4929)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4944..4965)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     misc_feature    5039..5875
                     /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(5916..5934)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(5941..5957)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     CDS             6095..6394
                     /label=ccdB
                     /note="CcdB, a bacterial toxin that poisons DNA gyrase"
     CDS             complement(7797..8654)
                     /label=AmpR
                     /note="beta-lactamase"
     rep_origin      8778..9366
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    9654..9675
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        9690..9720
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    9728..9744
                     /label=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     9752..9768
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        9789..9807
                     /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 cgagatgacc
      961 gagtacaagc ccacggtgcg cctcgccacc cgcgacgacg tccccagggc cgtacgcacc
     1021 ctcgccgccg cgttcgccga ctaccccgcc acgcgccaca ccgtcgatcc ggaccgccac
     1081 atcgagcggg tcaccgagct gcaagaactc ttcctcacgc gcgtcgggct cgacatcggc
     1141 aaggtgtggg tcgcggacga cggcgccgcg gtggcggtct ggaccacgcc ggagagcgtc
     1201 gaagcggggg cggtgttcgc cgagatcggc ccgcgcatgg ccgagttgag cggttcccgg
     1261 ctggccgcgc agcaacagat ggaaggcctc ctggcgccgc accggcccaa ggagcccgcg
     1321 tggttcctgg ccaccgtcgg cgtctcgccc gaccaccagg gcaagggtct gggcagcgcc
     1381 gtcgtgctcc ccggagtgga ggcggccgag cgcgccgggg tgcccgcctt cctggagacc
     1441 tccgcgcccc gcaacctccc cttctacgag cggctcggct tcaccgtcac cgccgacgtc
     1501 gaggtgcccg aaggaccgcg cacctggtgc atgacccgca agcccggtgc ctgatctaga
     1561 gggcccgttt aaacccgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     1621 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     1681 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     1741 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     1801 gcggtgggct ctatgggtct cgacattgat tattgactag ttattaatag taatcaatta
     1861 cggggtcatt agttcatagc ccatatatgg agttccgcgt tacataactt acggtaaatg
     1921 gcccgcctgg ctgaccgccc aacgaccccc gcccattgac gtcaataatg acgtatgttc
     1981 ccatagtaac gccaataggg actttccatt gacgtcaatg ggtggagtat ttacggtaaa
     2041 ctgcccactt ggcagtacat caagtgtatc atatgccaag tacgccccct attgacgtca
     2101 atgacggtaa atggcccgcc tggcattatg cccagtacat gaccttatgg gactttccta
     2161 cttggcagta catctacgta ttagtcatcg ctattaccat ggtcgaggtg agccccacgt
     2221 tctgcttcac tctccccatc tcccccccct ccccaccccc aattttgtat ttatttattt
     2281 tttaattatt ttgtgcagcg atgggggcgg gggggggggg ggggcgcgcg ccaggcgggg
     2341 cggggcgggg cgaggggcgg ggcggggcga ggcggagagg tgcggcggca gccaatcaga
     2401 gcggcgcgct ccgaaagttt ccttttatgg cgaggcggcg gcggcggcgg ccctataaaa
     2461 agcgaagcgc gcggcgggcg gggagtcgct gcgacgctgc cttcgccccg tgccccgctc
     2521 cgccgccgcc tcgcgccgcc cgccccggct ctgactgacc gcgttactcc cacaggtgag
     2581 cgggcgggac ggcccttctc ctccgggctg taattagcgc ttggtttaat gacggcttgt
     2641 ttcttttctg tggctgcgtg aaagccttga ggggctccgg gagggccctt tgtgcggggg
     2701 gagcggctcg gggggtgcgt gcgtgtgtgt gtgcgtgggg agcgccgcgt gcggctccgc
     2761 gctgcccggc ggctgtgagc gctgcgggcg cggcgcgggg ctttgtgcgc tccgcagtgt
     2821 gcgcgagggg agcgcggccg ggggcggtgc cccgcggtgc ggggggggct gcgaggggaa
     2881 caaaggctgc gtgcggggtg tgtgcgtggg ggggtgagca gggggtgtgg gcgcgtcggt
     2941 cgggctgcaa ccccccctgc acccccctcc ccgagttgct gagcacggcc cggcttcggg
     3001 tgcggggctc cgtacggggc gtggcgcggg gctcgccgtg ccgggcgggg ggtggcggca
     3061 ggtgggggtg ccgggcgggg cggggccgcc tcgggccggg gagggctcgg gggaggggcg
     3121 cggcggcccc cggagcgccg gcggctgtcg aggcgcggcg agccgcagcc attgcctttt
     3181 atggtaatcg tgcgagaggg cgcagggact tcctttgtcc caaatctgtg cggagccgaa
     3241 atctgggagg cgccgccgca ccccctctag cgggcgcggg gcgaagcggt gcggcgccgg
     3301 caggaaggaa atgggcgggg agggccttcg tgcgtcgccg cgccgccgtc cccttctccc
     3361 tctccagcct cggggctgtc cgcgggggga cggctgcctt cgggggggac ggggcagggc
     3421 ggggttcggc ttctggcgtg tgaccggcgg ctctagagcc tctgctaacc atgttcatgc
     3481 cttcttcttt ttcctacagc tcctgggcaa cgtgctggtt attgtgctgt ctcatcattt
     3541 tggcaaagaa ttccgccacc atggtgagca agggcgagga gctgttcacc ggggtggtgc
     3601 ccatcctggt cgagctggac ggcgacgtaa acggccacaa gttcagcgtg tccggcgagg
     3661 gcgagggcga tgccacctac ggcaagctga ccctgaagtt catctgcacc accggcaagc
     3721 tgcccgtgcc ctggcccacc ctcgtgacca ccctgaccta cggcgtgcag tgcttcagcc
     3781 gctaccccga ccacatgaag cagcacgact tcttcaagtc cgccatgccc gaaggctacg
     3841 tccaggagcg caccatcttc ttcaaggacg acggcaacta caagacccgc gccgaggtga
     3901 agttcgaggg cgacaccctg gtgaaccgca tcgagctgaa gggcatcgac ttcaaggagg
     3961 acggcaacat cctggggcac aagctggagt acaactacaa cagccacaac gtctatatca
     4021 tggccgacaa gcagaagaac ggcatcaagg tgaacttcaa gatccgccac aacatcgagg
     4081 acggcagcgt gcagctcgcc gaccactacc agcagaacac ccccatcggc gacggccccg
     4141 tgctgctgcc cgacaaccac tacctgagca cccagtccgc cctgagcaaa gaccccaacg
     4201 agaagcgcga tcacatggtc ctgctggagt tcgtgaccgc cgccgggatc actctcggca
     4261 tggacgagct gtacaagtaa acgcgtgaat tcactcctca ggtgcaggct gcctatcaga
     4321 aggtggtggc tggtgtggcc aatgccctgg ctcacaaata ccactgagat ctttttccct
     4381 ctgccaaaaa ttatggggac atcatgaagc cccttgagca tctgacttct ggctaataaa
     4441 ggaaatttat tttcattgca atagtgtgtt ggaatttttt gtgtctctca ctcggaagga
     4501 catatgggag ggcaaatcat ttaaaacatc agaatgagta tttggtttag agtttggcaa
     4561 catatgccca tatgctggct gccatgaaca aaggttggct ataaagaggt catcagtata
     4621 tgaaacagcc ccctgctgtc cattccttat tccatagaaa agccttgact tgaggttaga
     4681 ttttttttat attttgtttt gtgttatttt tttctttaac atccctaaaa ttttccttac
     4741 atgttttact agccagattt ttcctcctct cctgactact cccagtcata gctgtccctc
     4801 ttctcttatg gagatccctc gacctgcagc ccaagcttgg cgtaatcatg gtcatagctg
     4861 tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc cggaagcata
     4921 aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc gttgcgctca
     4981 ctgcccgctt tccagtcggg aaacctgtcg tgccagcgga tcgacagtac taagctttac
     5041 tagggacagg attggtgaca gaaaagcccc atccttaggc ctcctccttc ctagtctcct
     5101 gatattgggt ctaaccccca cctcctgtta ggcagattcc ttatctggtg acacaccccc
     5161 atttcctgga gccatctctc tccttgccag aacctctaag gtttgcttac gatggagcca
     5221 gagaggatcc tgggagggag agcttggcag ggggtgggag ggaagggggg gatgcgtgac
     5281 ctgcccggtt ctcagtggcc accctgcgct accctctccc agaacctgag ctgctctgac
     5341 gcggctgtct ggtgcgtttc actgatcctg gtgctgcagc ttccttacac ttcccaagag
     5401 gagaagcagt ttggaaaaac aaaatcagaa taagttggtc ctgagttcta actttggctc
     5461 ttcacctttc tagtccccaa tttatattgt tcctccgtgc gtcagtttta cctgtgagat
     5521 aaggccagta gccagccccg tcctggcagg gctgtggtga ggaggggggt gtccgtgtgg
     5581 aaaactccct ttgtgagaat ggtgcgtcct aggtgttcac caggtcgtgg ccgcctctac
     5641 tccctttctc tttctccatc cttctttcct taaagagtcc ccagtgctat ctgggacata
     5701 ttcctccgcc cagagcaggg tcccgcttcc ctaaggccct gctctgggct tctgggtttg
     5761 agtccttggc aagcccagga gaggcgctca ggcttccctg tcccccttcc tcgtccacca
     5821 tctcatgccc ctggctctcc tgccccttcc ctacaggggt tcctggctct gctctaaggg
     5881 caagggcgaa ttcgcggccg ctaaattcaa ttcgccctat agtgagtcgt attacaattc
     5941 actggccgtc gttttacaac gtcgtgactg ggaaaaccct ggcgttaccc aacttaatcg
     6001 ccttgcagca catccccctt tcgccagctg gcgtaatagc gaagaggccc gcaccgatcg
     6061 cccttcccaa cagttgcgca gcctatacgt acggcagttt aaggtttaca cctataaaag
     6121 agagagccgt tatcgtctgt ttgtggatgt acagagtgat attattgaca cgccggggcg
     6181 acggatggtg atccccctgg ccagtgcacg tctgctgtca gataaagtct cccgtgaact
     6241 ttacccggtg gtgcatatcg gggatgaaag ctggcgcatg atgaccaccg atatggccag
     6301 tgtgccggtc tccgttatcg gggaagaagt ggctgatctc agccaccgcg aaaatgacat
     6361 caaaaacgcc attaacctga tgttctgggg aatataaatg tcaggcatga gattatcaaa
     6421 aaggatcttc acctagatcc ttttcacgta gaaagccagt ccgcagaaac ggtgctgacc
     6481 ccggatgaat gtcagctact gggctatctg gacaagggaa aacgcaagcg caaagagaaa
     6541 gcaggtagct tgcagtgggc ttacatggcg atagctagac tgggcggttt tatggacagc
     6601 aagcgaaccg gaattgccag ctggggcgcc ctctggtaag gttgggaagc cctgcaaagt
     6661 aaactggatg gctttcttgc cgccaaggat ctgatggcgc aggggatcaa gctctgatca
     6721 agagacagga tgaggatcgt ttcgcatgat tgaacaagat ggattgcacg caggttctcc
     6781 ggccgcttgg gtggagaggc tattcggcta tgactgggca caacagacaa tcggctgctc
     6841 tgatgccgcc gtgttccggc tgtcagcgca ggggcgcccg gttctttttg tcaagaccga
     6901 cctgtccggt gccctgaatg aactgcaaga cgaggcagcg cggctatcgt ggctggccac
     6961 gacgggcgtt ccttgcgcag ctgtgctcga cgttgtcact gaagcgggaa gggactggct
     7021 gctattgggc gaagtgccgg ggcaggatct cctgtcatct caccttgctc ctgccgagaa
     7081 agtatccatc atggctgatg caatgcggcg gctgcatacg cttgatccgg ctacctgccc
     7141 attcgaccac caagcgaaac atcgcatcga gcgagcacgt actcggatgg aagccggtct
     7201 tgtcgatcag gatgatctgg acgaagagca tcaggggctc gcgccagccg aactgttcgc
     7261 caggctcaag gcgagcatgc ccgacggcga ggatctcgtc gtgacccatg catggcgatg
     7321 cctgcttgcc gaatatcatg gtggaaaatg gccgcttttc tggattcatc gactgtggcc
     7381 ggctgggtgt ggcggaccgc tatcaggaca tagcgttggc tacccgtgat attgctgaag
     7441 agcttggcgg cgaatgggct gaccgcttcc tcgtgcttta cggtatcgcc gctcccgatt
     7501 cgcagcgcat cgccttctat cgccttcttg acgagttctt ctgaattatt aacgcttaca
     7561 atttcctgat gcggtatttt ctccttacgc atctgtgcgg tatttcacac cgcatcaggt
     7621 ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca
     7681 aatatgtatc cgctcatgag attatcaaaa aggatcttca cctagatcct tttaaattaa
     7741 aaatgaagtt ttaaatcaat ctaaagtata tatgagtaaa cttggtctga cagttaccaa
     7801 tgcttaatca gtgaggcacc tatctcagcg atctgtctat ttcgttcatc catagttgcc
     7861 tgactccccg tcgtgtagat aactacgata cgggagggct taccatctgg ccccagtgct
     7921 gcaatgatac cgcgagaccc acgctcaccg gctccagatt tatcagcaat aaaccagcca
     7981 gccggaaggg ccgagcgcag aagtggtcct gcaactttat ccgcctccat ccagtctatt
     8041 aattgttgcc gggaagctag agtaagtagt tcgccagtta atagtttgcg caacgttgtt
     8101 gccattgcta caggcatcgt ggtgtcacgc tcgtcgtttg gtatggcttc attcagctcc
     8161 ggttcccaac gatcaaggcg agttacatga tcccccatgt tgtgcaaaaa agcggttagc
     8221 tccttcggtc ctccgatcgt tgtcagaagt aagttggccg cagtgttatc actcatggtt
     8281 atggcagcac tgcataattc tcttactgtc atgccatccg taagatgctt ttctgtgact
     8341 ggtgagtact caaccaagtc attctgagaa tagtgtatgc ggcgaccgag ttgctcttgc
     8401 ccggcgtcaa tacgggataa taccgcgcca catagcagaa ctttaaaagt gctcatcatt
     8461 ggaaaacgtt cttcggggcg aaaactctca aggatcttac cgctgttgag atccagttcg
     8521 atgtaaccca ctcgtgcacc caactgatct tcagcatctt ttactttcac cagcgtttct
     8581 gggtgagcaa aaacaggaag gcaaaatgcc gcaaaaaagg gaataagggc gacacggaaa
     8641 tgttgaatac tcatactctt cctttttcaa tattattgaa gcatttatca gggttattgt
     8701 ctcatgacca aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa
     8761 aagatcaaag gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca
     8821 aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt
     8881 ccgaaggtaa ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg
     8941 tagttaggcc accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc
     9001 ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga
     9061 cgatagttac cggataaggc gcagcggtcg ggctgaacgg ggggttcgtg cacacagccc
     9121 agcttggagc gaacgaccta caccgaactg agatacctac agcgtgagct atgagaaagc
     9181 gccacgcttc ccgaagggag aaaggcggac aggtatccgg taagcggcag ggtcggaaca
     9241 ggagagcgca cgagggagct tccaggggga aacgcctggt atctttatag tcctgtcggg
     9301 tttcgccacc tctgacttga gcgtcgattt ttgtgatgct cgtcaggggg gcggagccta
     9361 tggaaaaacg ccagcaacgc ggccttttta cggttcctgg ccttttgctg gccttttgct
     9421 cacatgttct ttcctgcgtt atcccctgat tctgtggata accgtattac cgcctttgag
     9481 tgagctgata ccgctcgccg cagccgaacg accgagcgca gcgagtcagt gagcgaggaa
     9541 gcggaagagc gcccaatacg caaaccgcct ctccccgcgc gttggccgat tcattaatgc
     9601 agctggcacg acaggtttcc cgactggaaa gcgggcagtg agcgcaacgc aattaatgtg
     9661 agttagctca ctcattaggc accccaggct ttacacttta tgcttccggc tcgtatgttg
     9721 tgtggaattg tgagcggata acaatttcac acaggaaaca gctatgacca tgattacgcc
     9781 aagctcagaa ttaaccctca ctaaaggga
//
