LOCUS       62056_2605        9190 bp DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9190)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..9190
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(1..105)
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     misc_feature    1402..2205
                     /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    2234..2575
                     /label=MESV Psi
                     /note="packaging signal of murine embryonic stem cell
                     virus"
     CDS             2642..3058
                     /codon_start=1
                     /label=gag (truncated)
                     /note="truncated Moloney murine leukemia virus (MMLV) gag
                     gene lacking the start codon"
                     /translation="GQTVTTPLSLTLGHWKDVERIAHNQSVDVKKRRWVTFCSAEWPTF
                     NVGWPRDGTFNRDLITQVKIKVFSPGPHGHPDQVPYIVTWEALAFDPPPWVKPFVHPKP
                     PPPLPPSAPSLPLEPPRSTPPRSSLYPALTPSLGA"
     polyA_signal    3098..3322
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     promoter        3369..3580
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             3593..3861
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             3917..4582
                     /codon_start=1
                     /label=CopGFP
                     /note="green fluorescent protein 2 from Pontellina plumata,
                     also known as ppluGFP2 (Shagin et al., 2004)"
                     /translation="LPAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             4652..4705
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             4706..5302
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    5413..5545
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    5798..6634
                     /label=HA-R
                     /note="right homology arm from the adeno-associated virus 
                     integration site (AAVS1) within intron 1 of the human 
                     PPP1R12C gene"
     primer_bind     complement(7169..7185)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(7193..7209)
                     /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(7217..7247)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(7262..7283)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(7571..8159)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(8333..9190)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtaacggc cgccagtgtg
     1381 ctggaattcg cccttggtac ctgctttctc tgaccagcat tctctcccct gggcctgtgc
     1441 cgctttctgt ctgcagcttg tggcctgggt cacctctacg gctggcccag atccttccct
     1501 gccgcctcct tcaggttccg tcttcctcca ctccctcttc cccttgctct ctgctgtgtt
     1561 gctgcccaag gatgctcttt ccggagcact tccttctcgg cgctgcacca cgtgatgtcc
     1621 tctgagcgga tcctccccgt gtctgggtcc tctccgggca tctctcctcc ctcacccaac
     1681 cccatgccgt cttcactcgc tgggttccct tttccttctc cttctggggc ctgtgccatc
     1741 tctcgtttct taggatggcc ttctccgacg gatgtctccc ttgcgtcccg cctccccttc
     1801 ttgtaggcct gcatcatcac cgtttttctg gacaacccca aagtaccccg tctccctggc
     1861 tttagccacc tctccatcct cttgctttct ttgcctggac accccgttct cctgtggatt
     1921 cgggtcacct ctcactcctt tcatttgggc agctccccta ccccccttac ctctctagtc
     1981 tgtgctagct cttccagccc cctgtcatgg catcttccag gggtccgaga gctcagctag
     2041 tcttcttcct ccaacccggg cccctatgtc cacttcagga cagcatgttt gctgcctcca
     2101 gggatcctgt gtccccgagc tgggaccacc ttatattccc agggccggtt aatgtggctc
     2161 tggttctggg tacttttatc tgtcccctcc accccacagt ggggcacgcg taagggcgaa
     2221 ttctgcagat atcaagctgg ccagcggtcg tttcgtgtct gtctctgtct ttgtgcgtgt
     2281 ttgtgccggc atctaatgtt tgcgcctgcg tctgtactag ttagctaact agctctgtat
     2341 ctggcggacc cgtggtggaa ctgacgagtt ctgaacaccc ggccgcaacc ctgggagacg
     2401 tcccagggac tttgggggcc gtttttgtgg cccgacctga ggaagggagt cgatgtggaa
     2461 tccgaccccg tcaggatatg tggttctggt aggagacgag aacctaaaac agttcccgcc
     2521 tccgtctgaa tttttgcttt cggtttggaa ccgaagccgc gcgtcttgtc tgctgcagcg
     2581 ctgcagcatc gttctgtgtt gtctctgtct gactgtgttt ctgtatttgt ctgaaaatta
     2641 gggccagact gttaccactc ccttaagttt gaccttaggt cactggaaag atgtcgagcg
     2701 gatcgctcac aaccagtcgg tagatgtcaa gaagagacgt tgggttacct tctgctctgc
     2761 agaatggcca acctttaacg tcggatggcc gcgagacggc acctttaacc gagacctcat
     2821 cacccaggtt aagatcaagg tcttttcacc tggcccgcat ggacacccag accaggtccc
     2881 ctacatcgtg acctgggaag ccttggcttt tgacccccct ccctgggtca agccctttgt
     2941 acaccctaag cctccgcctc ctcttcctcc atccgccccg tctctccccc ttgaacctcc
     3001 tcgttcgacc ccgcctcgat cctcccttta tccagccctc actccttctc taggcgccgg
     3061 aattaccggt cacgtgattc gaatttaaat cggatccctg tgccttctag ttgccagcca
     3121 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3181 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3241 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3301 ggggatgcgg tgggctctat ggagatctgc ggccgcaagg atctgcgatc gctccggtgc
     3361 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     3421 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     3481 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     3541 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     3601 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     3661 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     3721 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     3781 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     3841 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga
     3901 gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa
     3961 cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac
     4021 caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt
     4081 gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct
     4141 gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg
     4201 cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa
     4261 ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag
     4321 caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc
     4381 ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca
     4441 cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg
     4501 ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt
     4561 caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg
     4621 caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg
     4681 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     4741 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     4801 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     4861 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     4921 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     4981 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     5041 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     5101 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     5161 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     5221 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     5281 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt
     5341 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     5401 tttgtatcag ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     5461 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     5521 atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt ctatcagaag
     5581 ctcatctggt ctcccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt
     5641 tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc
     5701 tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg gaattgactg
     5761 gattcctttt ttagggccca ttggtatggt gtacactact agggacagga ttggtgacag
     5821 aaaagcccca tccttaggcc tcctccttcc tagtctcctg atattgggtc taacccccac
     5881 ctcctgttag gcagattcct tatctggtga cacaccccca tttcctggag ccatctctct
     5941 ccttgccaga acctctaagg tttgcttacg atggagccag agaggatcct gggagggaga
     6001 gcttggcagg gggtgggagg gaaggggggg atgcgtgacc tgcccggttc tcagtggcca
     6061 ccctgcgcta ccctctccca gaacctgagc tgctctgacg cggctgtctg gtgcgtttca
     6121 ctgatcctgg tgctgcagct tccttacact tcccaagagg agaagcagtt tggaaaaaca
     6181 aaatcagaat aagttggtcc tgagttctaa ctttggctct tcacctttct agtccccaat
     6241 ttatattgtt cctccgtgcg tcagttttac ctgtgagata aggccagtag ccagccccgt
     6301 cctggcaggg ctgtggtgag gaggggggtg tccgtgtgga aaactccctt tgtgagaatg
     6361 gtgcgtccta ggtgttcacc aggtcgtggc cgcctctact ccctttctct ttctccatcc
     6421 ttctttcctt aaagagtccc cagtgctatc tgggacatat tcctccgccc agagcagggt
     6481 cccgcttccc taaggccctg ctctgggctt ctgggtttga gtccttggca agcccaggag
     6541 aggcgctcag gcttccctgt cccccttcct cgtccaccat ctcatgcccc tggctctcct
     6601 gccccttccc tacaggtgtt cctggctctg ctctctcgag atgcatgcgt caattttacg
     6661 cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc
     6721 cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc
     6781 ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt
     6841 gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc
     6901 tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg
     6961 acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc
     7021 cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc
     7081 agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta
     7141 cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     7201 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     7261 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     7321 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     7381 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     7441 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     7501 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     7561 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     7621 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     7681 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     7741 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     7801 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     7861 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     7921 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     7981 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     8041 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     8101 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     8161 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     8221 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     8281 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     8341 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     8401 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     8461 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     8521 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     8581 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     8641 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     8701 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     8761 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     8821 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     8881 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     8941 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     9001 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     9061 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     9121 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     9181 gaatactcat
//
