LOCUS       62056_2600        8110 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 8110)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..8110
                     /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"
     promoter        2289..2500
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             2513..2781
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2837..3502
                     /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             3572..3625
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3626..4222
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4333..4465
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     misc_feature    4718..5554
                     /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(6089..6105)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(6113..6129)
                     /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(6137..6167)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(6182..6203)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6491..7079)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(7253..8110)
                     /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 atcactgaga tctactggcg gccgcgaagg atctgcgatc gctccggtgc
     2281 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     2341 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     2401 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     2461 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     2521 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     2581 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     2641 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     2701 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     2761 tgttctgcgc cgttacagat ccaagctgtg accggcgcct acgctagacg ccaccatgga
     2821 gagcgacgag agcggcctgc ccgccatgga gatcgagtgc cgcatcaccg gcaccctgaa
     2881 cggcgtggag ttcgagctgg tgggcggcgg agagggcacc cccaagcagg gccgcatgac
     2941 caacaagatg aagagcacca aaggcgccct gaccttcagc ccctacctgc tgagccacgt
     3001 gatgggctac ggcttctacc acttcggcac ctaccccagc ggctacgaga accccttcct
     3061 gcacgccatc aacaacggcg gctacaccaa cacccgcatc gagaagtacg aggacggcgg
     3121 cgtgctgcac gtgagcttca gctaccgcta cgaggccggc cgcgtgatcg gcgacttcaa
     3181 ggtggtgggc accggcttcc ccgaggacag cgtgatcttc accgacaaga tcatccgcag
     3241 caacgccacc gtggagcacc tgcaccccat gggcgataac gtgctggtgg gcagcttcgc
     3301 ccgcaccttc agcctgcgcg acggcggcta ctacagcttc gtggtggaca gccacatgca
     3361 cttcaagagc gccatccacc ccagcatcct gcagaacggg ggccccatgt tcgccttccg
     3421 ccgcgtggag gagctgcaca gcaacaccga gctgggcatc gtggagtacc agcacgcctt
     3481 caagaccccc atcgccttcg ccagatcccg cgctcagtcg tccaattctg ccgtggacgg
     3541 caccgccgga cccggctcca ccggatctcg cgagggcaga ggaagtcttc taacatgcgg
     3601 tgacgtggag gagaatcccg gccctatgac cgagtacaag cccacggtgc gcctcgccac
     3661 ccgcgacgac gtccccaggg ccgtacgcac cctcgccgcc gcgttcgccg actaccccgc
     3721 cacgcgccac accgtcgatc cggaccgcca catcgagcgg gtcaccgagc tgcaagaact
     3781 cttcctcacg cgcgtcgggc tcgacatcgg caaggtgtgg gtcgcggacg acggcgccgc
     3841 ggtggcggtc tggaccacgc cggagagcgt cgaagcgggg gcggtgttcg ccgagatcgg
     3901 cccgcgcatg gccgagttga gcggttcccg gctggccgcg cagcaacaga tggaaggcct
     3961 cctggcgccg caccggccca aggagcccgc gtggttcctg gccaccgtcg gcgtctcgcc
     4021 cgaccaccag ggcaagggtc tgggcagcgc cgtcgtgctc cccggagtgg aggcggccga
     4081 gcgcgccggg gtgcccgcct tcctggagac ctccgcgccc cgcaacctcc ccttctacga
     4141 gcggctcggc ttcaccgtca ccgccgacgt cgaggtgccc gaaggaccgc gcacctggtg
     4201 catgacccgc aagcccggtg cctgaaatca acctctggat tacaaaattt gtgaaagatt
     4261 gactggtatt cttaactatg ttgctccttt tacgctatgt ggatacgctg ctttaatgcc
     4321 tttgtatcag ttaacttgtt tattgcagct tataatggtt acaaataaag caatagcatc
     4381 acaaatttca caaataaagc atttttttca ctgcattcta gttgtggttt gtccaaactc
     4441 atcaatgtat cttatcatgt ctggaattga ctcaaatgat gtcaattagt ctatcagaag
     4501 ctcatctggt ctcccttccg ggggacaaga catccctgtt taatatttaa acagcagtgt
     4561 tcccaaactg ggttcttata tcccttgctc tggtcaacca ggttgcaggg tttcctgtcc
     4621 tcacaggaac gaagtcccta aagaaacagt ggcagccagg tttagccccg gaattgactg
     4681 gattcctttt ttagggccca ttggtatggt gtacactact agggacagga ttggtgacag
     4741 aaaagcccca tccttaggcc tcctccttcc tagtctcctg atattgggtc taacccccac
     4801 ctcctgttag gcagattcct tatctggtga cacaccccca tttcctggag ccatctctct
     4861 ccttgccaga acctctaagg tttgcttacg atggagccag agaggatcct gggagggaga
     4921 gcttggcagg gggtgggagg gaaggggggg atgcgtgacc tgcccggttc tcagtggcca
     4981 ccctgcgcta ccctctccca gaacctgagc tgctctgacg cggctgtctg gtgcgtttca
     5041 ctgatcctgg tgctgcagct tccttacact tcccaagagg agaagcagtt tggaaaaaca
     5101 aaatcagaat aagttggtcc tgagttctaa ctttggctct tcacctttct agtccccaat
     5161 ttatattgtt cctccgtgcg tcagttttac ctgtgagata aggccagtag ccagccccgt
     5221 cctggcaggg ctgtggtgag gaggggggtg tccgtgtgga aaactccctt tgtgagaatg
     5281 gtgcgtccta ggtgttcacc aggtcgtggc cgcctctact ccctttctct ttctccatcc
     5341 ttctttcctt aaagagtccc cagtgctatc tgggacatat tcctccgccc agagcagggt
     5401 cccgcttccc taaggccctg ctctgggctt ctgggtttga gtccttggca agcccaggag
     5461 aggcgctcag gcttccctgt cccccttcct cgtccaccat ctcatgcccc tggctctcct
     5521 gccccttccc tacaggtgtt cctggctctg ctctctcgag atgcatgcgt caattttacg
     5581 cagactatct ttctagggtt aatctagctg catcaggatc atatcgtcgg gtcttttttc
     5641 cggctcagtc atcgcccaag ctggcgctat ctgggcatcg gggaggaaga agcccgtgcc
     5701 ttttcccgcg aggttgaagc ggcatggaaa gagtttgccg aggatgactg ctgctgcatt
     5761 gacgttgagc gaaaacgcac gtttaccatg atgattcggg aaggtgtggc catgcacgcc
     5821 tttaacggtg aactgttcgt tcaggccacc tgggatacca gttcgtcgcg gcttttccgg
     5881 acacagttcc ggatggtcag cccgaagcgc atcagcaacc cgaacaatac cggcgacagc
     5941 cggaactgcc gtgccggtgt gcagattaat gacagcggtg cggcgctggg atattacgtc
     6001 agcgaggacg ggtatcctgg ctggatgccg cagaaatgga catggatacc ccgtgagtta
     6061 cccggcgggc gcgcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     6121 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     6181 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     6241 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     6301 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     6361 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     6421 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     6481 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     6541 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     6601 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     6661 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     6721 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     6781 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     6841 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     6901 agtggtggcc taactacggc tacactagaa ggacagtatt tggtatctgc gctctgctga
     6961 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     7021 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     7081 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     7141 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     7201 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     7261 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     7321 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     7381 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     7441 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     7501 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     7561 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     7621 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     7681 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     7741 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     7801 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     7861 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     7921 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     7981 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     8041 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     8101 gaatactcat
//
