LOCUS       40924_40382        6486 bp DNA     circular SYN 08-APR-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6486)
  AUTHORS   caoheibi
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6486)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6486
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     LTR             733..1249
                     /label=5' LTR
                     /note="5' long terminal repeat from murine embryonic stem
                     cell virus"
     misc_feature    1313..1654
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             1721..2137
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     primer_bind     2156..2172
                     /label=KS primer
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     promoter        complement(2246..2486)
                     /label=U6 promoter
                     /note="RNA polymerase III promoter for human U6 snRNA"
     promoter        2580..3079
                     /label=PGK promoter
                     /note="mouse phosphoglycerate kinase 1 promoter"
     CDS             3104..3700
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     protein_bind    complement(4289..4305)
                     /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(4313..4343)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(4358..4379)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4667..5255)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5429..6286)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     promoter        complement(6287..6391)
                     /label=AmpR promoter
ORIGIN
        1 tcgcgcgttt cggtgatgac ggtgaaaacc tctgacacat gcagctcccg gagacggtca
       61 cagcttgtct gtaagcggat gccgggagca gacaagcccg tcagggcgcg tcagcgggtg
      121 ttggcgggtg tcggggctgg cttaactatg cggcatcaga gcagattgta ctgagagtgc
      181 accatatgcg gtgtgaaata ccgcacagat gcgtaaggag aaaataccgc atcaggcgcc
      241 attcgccatt caggctgcgc aactgttggg aagggcgatc ggtgcgggcc tcttcgctat
      301 tacgccagct ggcgaaaggg ggatgtgctg caaggcgatt aagttgggta acgccagggt
      361 tttcccagtc acgacgttgt aaaacgacgg cgcaaggaat ggtgcatgca aggagatggc
      421 gcccaacagt cccccggcca cggggcctgc caccataccc acgccgaaac aagcgctcat
      481 gagcccgaag tggcgagccc gatcttcccc atcggtgatg tcggcgatat aggcgccagc
      541 aaccgcacct gtggcgccgg tgatgccggc cacgatgcgt ccggcgtaga ggcgattagt
      601 ccaatttgtt aaagacagga tatcagtggt ccaggctcta gttttgactc aacaatatca
      661 ccagctgaag cctatagagt acgagccata gataaaataa aagattttat ttagtctcca
      721 gaaaaagggg ggaatgaaag accccacctg taggtttggc aagctagctt aagtaacgcc
      781 attttgcaag gcatggaaaa tacataactg agaatagaga agttcagatc aaggttagga
      841 acagagagac agcagaatat gggccaaaca ggatatctgt ggtaagcagt tcctgccccg
      901 gctcagggcc aagaacagat ggtccccaga tgcggtcccg ccctcagcag tttctagaga
      961 accatcagat gtttccaggg tgccccaagg acctgaaatg accctgtgcc ttatttgaac
     1021 taaccaatca gttcgcttct cgcttctgtt cgcgcgcttc tgctccccga gctcaataaa
     1081 agagcccaca acccctcact cggcgcgcca gtcctccgat agactgcgtc gcccgggtac
     1141 ccgtattccc aataaagcct cttgctgttt gcatccgaat cgtggactcg ctgatccttg
     1201 ggagggtctc ctcagattga ttgactgccc acctcggggg tctttcattt ggaggttcca
     1261 ccgagatttg gagacccctg cccagggacc accgaccccc ccgccgggag gtaagctggc
     1321 cagcggtcgt ttcgtgtctg tctctgtctt tgtgcgtgtt tgtgccggca tctaatgttt
     1381 gcgcctgcgt ctgtactagt tagctaacta gctctgtatc tggcggaccc gtggtggaac
     1441 tgacgagttc tgaacacccg gccgcaaccc tgggagacgt cccagggact ttgggggccg
     1501 tttttgtggc ccgacctgag gaagggagtc gatgtggaat ccgaccccgt caggatatgt
     1561 ggttctggta ggagacgaga acctaaaaca gttcccgcct ccgtctgaat ttttgctttc
     1621 ggtttggaac cgaagccgcg cgtcttgtct gctgcagcgc tgcagcatcg ttctgtgttg
     1681 tctctgtctg actgtgtttc tgtatttgtc tgaaaattag ggccagactg ttaccactcc
     1741 cttaagtttg accttaggtc actggaaaga tgtcgagcgg atcgctcaca accagtcggt
     1801 agatgtcaag aagagacgtt gggttacctt ctgctctgca gaatggccaa cctttaacgt
     1861 cggatggccg cgagacggca cctttaaccg agacctcatc acccaggtta agatcaaggt
     1921 cttttcacct ggcccgcatg gacacccaga ccaggtcccc tacatcgtga cctgggaagc
     1981 cttggctttt gacccccctc cctgggtcaa gccctttgta caccctaagc ctccgcctcc
     2041 tcttcctcca tccgccccgt ctctccccct tgaacctcct cgttcgaccc cgcctcgatc
     2101 ctccctttat ccagccctca ctccttctct aggcgccgga attagatcga tctctcgagg
     2161 tcgacggtat cgataagctt ttccaaaaaa gtactgctta cgatacggtc tcttgaaccg
     2221 tatcgtaagc agtactggat cccgcgtcct ttccacaaga tatataaacc caagaaatcg
     2281 aaatactttc aagttacggt aagcatatga tagtccattt taaaacataa ttttaaaact
     2341 gcaaactacc caagaaatta ttactttcta cgtcacgtat tttgtactaa tatctttgtg
     2401 tttacagtca aattaattct aattatctct ctaacagcct tgtatcgtat atgcaaatat
     2461 gaaggaatca tgggaaatag gccctcttcc tgcccgacct tggcgcgcgc tcggcgcgcg
     2521 gtcacgctcc gtcacgtggt gcgttttgcc tgcgcgtctt tccactgggg aattctaccg
     2581 ggtaggggag gcgcttttcc caaggcagtc tggagcatgc gctttagcag ccccgctggg
     2641 cacttggcgc tacacaagtg gcctctggcc tcgcacacat tccacatcca ccggtaggcg
     2701 ccaaccggct ccgttctttg gtggcccctt cgcgccacct tctactcctc ccctagtcag
     2761 gaagttcccc cccgccccgc agctcgcgtc gtgcaggacg tgacaaatgg aagtagcacg
     2821 tctcactagt ctcgtgcaga tggacagcac cgctgagcaa tggaagcggg taggcctttg
     2881 gggcagcggc caatagcagc tttgctcctt cgctttctgg gctcagaggc tgggaagggg
     2941 tgggtccggg ggcgggctca ggggcgggct caggggcggg gcgggcgccc gaaggtcctc
     3001 cggaggcccg gcattctgca cgcttcaaaa gcgcacgtct gccgcgctgt tctcctcttc
     3061 ctcatctccg ggcctttcga cctgcagccc aagctagctt accatgaccg agtacaagcc
     3121 cacggtgcgc ctcgccaccc gcgacgacgt ccccagggcc gtacgcaccc tcgccgccgc
     3181 gttcgccgac taccccgcca cgcgccacac cgtcgatccg gaccgccaca tcgagcgggt
     3241 caccgagctg caagaactct tcctcacgcg cgtcgggctc gacatcggca aggtgtgggt
     3301 cgcggacgac ggcgccgcgg tggcggtctg gaccacgccg gagagcgtcg aagcgggggc
     3361 ggtgttcgcc gagatcggcc cgcgcatggc cgagttgagc ggttcccggc tggccgcgca
     3421 gcaacagatg gaaggcctcc tggcgccgca ccggcccaag gagcccgcgt ggttcctggc
     3481 caccgtcggc gtctcgcccg accaccaggg caagggtctg ggcagcgccg tcgtgctccc
     3541 cggagtggag gcggccgagc gcgccggggt gcccgccttc ctggagacct ccgcgccccg
     3601 caacctcccc ttctacgagc ggctcggctt caccgtcacc gccgacgtcg aggtgcccga
     3661 aggaccgcgc acctggtgca tgacccgcaa gcccggtgcc tgacgcccgc cccacgaccc
     3721 gcagcgcccg accgaaagga gcgcacgacc ccatgcatcg ataaaataaa agattttatt
     3781 tagtctccag aaaaaggggg gaatgaaaga ccccacctgt aggtttggca agctagagaa
     3841 ccatcagatg tttccagggt gccccaagga cctgaaatga ccctgtgcct tatttgaact
     3901 aaccaatcag ttcgcttctc gcttctgttc gcgcgcttct gctccccgag ctcaataaaa
     3961 gagcccacaa cccctcactc ggcgcgccag tcctccgata gactgcgtcg cccgggtacc
     4021 cgtgtatcca ataaaccctc ttgcagttgc atccgacttg tggtctcgct gttccttggg
     4081 agggtctcct ctgagtgatt gactacccgt cagcgggggt ctttcatggg taacagtttc
     4141 ttgaagttgg agaacaacat tctgagggta ggagtcgaat attaagtaat cctgactcaa
     4201 ttagccactg ttttgaatcc acatactcca atactcctga aatagttcat tatggacagc
     4261 gcagaaagag ctggggagaa ttgtgaaatt gttatccgct cacaattcca cacaacatac
     4321 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4381 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4441 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4501 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4561 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4621 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4681 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4741 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4801 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4861 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4921 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4981 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     5041 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5101 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5161 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5221 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5281 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5341 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5401 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5461 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5521 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5581 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5641 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5701 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5761 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5821 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5881 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5941 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     6001 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6061 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6121 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6181 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6241 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6301 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6361 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6421 tctaagaaac cattattatc atgacattaa cctataaaaa taggcgtatc acgaggccct
     6481 ttcgtc
//
