LOCUS       V023384                 9516 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V023384
VERSION     V023384
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     promoter        complement(1..105)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     primer_bind     728..744
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     promoter        1442..1645
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             1734..3749
                     /label="SHCBP1(NM_024745)"
                     /note="SHCBP1(NM_024745)"
                     /gene="SHCBP1"
     polyA_signal    3778..4002
                     /note="bovine growth hormone polyadenylation signal"
                     /label="bGH poly(A) signal"
     promoter        4043..4254
                     /note="core&nbsp;promoter for human elongation factor
                     EF-1α"
                     /label="EF-1α core promoter"
     LTR             4267..4535
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
                     /label="5' LTR (truncated)"
     regulatory      4564..4573
                     /note="vertebrate consensus sequence for strong initiation
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
                     /label="Kozak sequence"
     CDS             4594..5256
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /transl_table=1
                     /translation="PAMEIECRITGTLNGVEFELVGGGEGTPKQGRMTNKMKSTKGALT
                     FSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYRY
                     EAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDGG
                     YYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA"
                     /label="CopGFP"
     CDS             5326..5379
                     /codon_start=1
                     /note="Eukaryotic ribosomes fail to insert a peptide bond
                     between the Gly and Pro residues, yielding separate
                     polypeptides."
                     /product="2A peptide from&nbsp;Thosea asigna virus capsid
                     protein"
                     /protein_id=""
                     /transl_table=1
                     /translation="EGRGSLLTCGDVEENPGP"
                     /label="T2A"
     CDS             5380..5979
                     /codon_start=1
                     /gene="pac from Streptomyces alboniger"
                     /note="confers resistance to puromycin"
                     /product="puromycin N-acetyltransferase"
                     /transl_table=1
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA*"
                     /label="PuroR"
     polyA_signal    6087..6208
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(7495..7511)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    7519..7535
                     /bound_moiety="lac repressor encoded by lacI"
                     /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-β-D-thiogalactopyranoside (IPTG)."
                     /label="lac operator"
     promoter        complement(7543..7573)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    7588..7609
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(7897..8485)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(8656..9516)
                     /codon_start=1
                     /gene="bla"
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /product="β-lactamase"
                     /transl_table=1
                     /translation="MSIQHFRVALIPFFAAFCLPVFA,HPETLVKVKDAEDQLGARVGY
                     IELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEY
                     SPVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDR
                     WEPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRS
                     ALPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGA
                     SLIKHW*"
                     /label="AmpR"
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 tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1501 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1561 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1621 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1681 tccacgctgt tttgacctcc atagaagatt ctagagctag cgaattcgcc accatggctg
     1741 acgggtcgct gacgggcggc ggtctggagg cagcggccat ggcgccggag cgcacgggct
     1801 gggcggtgga gcaggagctg gcgtctctgg agaaaggttt gttccaagat gaagattcat
     1861 gcagtgattg tagctaccgt gataaaccag gttctagttt acaaagtttt atgccagaag
     1921 gaaaaacctt tttcccagaa attttccaaa caaatcaact tttgttctat gagcgattca
     1981 gagcctatca agattacatt ttagctgact gcaaggcctc tgaggtacag gaattcacag
     2041 ctgagttctt ggagaaggtc cttgagccat ctggatggcg ggcagtctgg cacactaatg
     2101 tgttcaaggt gctggttgag atcacagatg tggactttgc agccttgaag gcagtggtga
     2161 ggcttgctga accatacctc tgtgactctc aagtgagcac ttttaccatg gagtgcatga
     2221 aggagctcct tgatctgaag gagcatcggt tgcccctgca ggagctgtgg gtggtgtttg
     2281 atgattcagg agtgtttgac cagacagccc ttgcaattga gcatgtcaga tttttctacc
     2341 aaaacatttg gaggagttgg gatgaagaag aggaggatga atacgattat tttgtcagat
     2401 gtgttgaacc tcgattaaga ttgcattatg acattcttga agaccgagtt ccatcaggac
     2461 ttattgttga ctaccacaat ctgttgtctc aatgtgagga gagttacagg aaatttttaa
     2521 atctgagaag cagtttgtca aattgtaact ctgattccga gcaggaaaat atctccatgg
     2581 tggaagggtt aaaattgtat tcggagatgg aacagttgaa acaaaagctg aaactcattg
     2641 agaatccttt gttgaggtat gtgtttggtt atcagaagaa ttctaacatc caagcaaagg
     2701 gtgtccgttc cagcggtcag aagatcactc atgtggtctc ctccaccatg atggctggtc
     2761 tcctgcggtc cctgcttacg gacaggcttt gccaggagcc tggtgaggaa gaaagagaaa
     2821 ttcagttcca tagtgatcca ttgtctgcta taaatgcctg cttcgaaggt gacactgtta
     2881 ttgtttgtcc tggccattat gtggtacatg gcactttctc cattgctgac tccattgagt
     2941 tggaaggata tggcctacca gatgacattg tgatagaaaa gaggggcaaa ggcgacactt
     3001 ttgtggactg cactggtgct gatattaaaa tctcaggcat aaaatttgtt cagcatgatg
     3061 ctgtagaggg aatcttaatt gttcaccgtg gtaagactac gctggaaaac tgtgtgctgc
     3121 agtgtgagac gaccggagtc acagtgcgga catcagcaga gtttctaatg aagaactcgg
     3181 atttatatgg cgccaagggt gctggtatag aaatctaccc tgggagtcag tgcaccctga
     3241 gtgacaatgg gatccatcac tgcaaggaag ggatcctcat taaggacttc ttagatgaac
     3301 attatgacat tcccaagata tccatggtga ataatataat acataataat gaaggttatg
     3361 gtgttgtctt ggtgaaacct acaatcttct ctgacctgca agaaaatgct gaagatggaa
     3421 ctgaagaaaa taaagcgctt aaaattcaga caagtggaga gccagatgtg gctgaaagag
     3481 tggatctaga agagctgatt gagtgtgcaa ctggtaaaat ggagctttgt gcaagaactg
     3541 acccttctga gcaagtcgag ggaaattgtg aaattgtaaa tgaactaatt gctgcctcca
     3601 cacagaaagg ccagataaag aagaaaaggt tgagtgaact ggggatcacg caagctgatg
     3661 acaacttaat gtcacaggag atgtttgttg ggattgtggg gaaccagttc aagtggaatg
     3721 ggaaaggtag ttttggcaca tttcttttct gagtttaaac ccgctgatca gcctcgactg
     3781 tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg
     3841 aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg cattgtctga
     3901 gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg gaggattggg
     3961 aagacaatag caggcatgct ggggatgcgg tgggctctat gggcggccgc aaggatctgc
     4021 gatcgctccg gtgcccgtca gtgggcagag cgcacatcgc ccacagtccc cgagaagttg
     4081 gggggagggg tcggcaattg aacgggtgcc tagagaaggt ggcgcggggt aaactgggaa
     4141 agtgatgtcg tgtactggct ccgccttttt cccgagggtg ggggagaacc gtatataagt
     4201 gcagtagtcg ccgtgaacgt tctttttcgc aacgggtttg ccgccagaac acagctgaag
     4261 cttcgagggg ctcgcatctc tccttcacgc gcccgccgcc ctacctgagg ccgccatcca
     4321 cgccggttga gtcgcgttct gccgcctccc gcctgtggtg cctcctgaac tgcgtccgcc
     4381 gtctaggtaa gtttaaagct caggtcgaga ccgggccttt gtccggcgct cccttggagc
     4441 ctacctagac tcagccggct ctccacgctt tgcctgaccc tgcttgctca actctacgtc
     4501 tttgtttcgt tttctgttct gcgccgttac agatccaagc tgtgaccggc gcctacgcta
     4561 gacgccacca tggagagcga cgagagcggc ctgcccgcca tggagatcga gtgccgcatc
     4621 accggcaccc tgaacggcgt ggagttcgag ctggtgggcg gcggagaggg cacccccaag
     4681 cagggccgca tgaccaacaa gatgaagagc accaaaggcg ccctgacctt cagcccctac
     4741 ctgctgagcc acgtgatggg ctacggcttc taccacttcg gcacctaccc cagcggctac
     4801 gagaacccct tcctgcacgc catcaacaac ggcggctaca ccaacacccg catcgagaag
     4861 tacgaggacg gcggcgtgct gcacgtgagc ttcagctacc gctacgaggc cggccgcgtg
     4921 atcggcgact tcaaggtggt gggcaccggc ttccccgagg acagcgtgat cttcaccgac
     4981 aagatcatcc gcagcaacgc caccgtggag cacctgcacc ccatgggcga taacgtgctg
     5041 gtgggcagct tcgcccgcac cttcagcctg cgcgacggcg gctactacag cttcgtggtg
     5101 gacagccaca tgcacttcaa gagcgccatc caccccagca tcctgcagaa cgggggcccc
     5161 atgttcgcct tccgccgcgt ggaggagctg cacagcaaca ccgagctggg catcgtggag
     5221 taccagcacg ccttcaagac ccccatcgcc ttcgccagat cccgcgctca gtcgtccaat
     5281 tctgccgtgg acggcaccgc cggacccggc tccaccggat ctcgcgaggg cagaggaagt
     5341 cttctaacat gcggtgacgt ggaggagaat cccggcccta tgaccgagta caagcccacg
     5401 gtgcgcctcg ccacccgcga cgacgtcccc agggccgtac gcaccctcgc cgccgcgttc
     5461 gccgactacc ccgccacgcg ccacaccgtc gatccggacc gccacatcga gcgggtcacc
     5521 gagctgcaag aactcttcct cacgcgcgtc gggctcgaca tcggcaaggt gtgggtcgcg
     5581 gacgacggcg ccgcggtggc ggtctggacc acgccggaga gcgtcgaagc gggggcggtg
     5641 ttcgccgaga tcggcccgcg catggccgag ttgagcggtt cccggctggc cgcgcagcaa
     5701 cagatggaag gcctcctggc gccgcaccgg cccaaggagc ccgcgtggtt cctggccacc
     5761 gtcggcgtct cgcccgacca ccagggcaag ggtctgggca gcgccgtcgt gctccccgga
     5821 gtggaggcgg ccgagcgcgc cggggtgccc gccttcctgg agacctccgc gccccgcaac
     5881 ctccccttct acgagcggct cggcttcacc gtcaccgccg acgtcgaggt gcccgaagga
     5941 ccgcgcacct ggtgcatgac ccgcaagccc ggtgcctgaa atcaacctct ggattacaaa
     6001 atttgtgaaa gattgactgg tattcttaac tatgttgctc cttttacgct atgtggatac
     6061 gctgctttaa tgcctttgta tcagttaact tgtttattgc agcttataat ggttacaaat
     6121 aaagcaatag catcacaaat ttcacaaata aagcattttt ttcactgcat tctagttgtg
     6181 gtttgtccaa actcatcaat gtatcttatc atgtctggaa ttgactcaaa tgatgtcaat
     6241 tagtctatca gaagctatct ggtctccctt ccgggggaca agacatccct gtttaatatt
     6301 taaacagcag tgttcccaaa ctgggttctt atatcccttg ctctggtcaa ccaggttgca
     6361 gggtttcctg tcctcacagg aacgaagtcc ctaaagaaac agtggcagcc aggtttagcc
     6421 ccggaattga ctggattcct tttttagggc ccattggtat ggctttttcc ccgtatcccc
     6481 ccaggtgtct gcaggctcaa agagcagcga gaagcgttca gaggaaagcg atcccgtgcc
     6541 accttccccg tgcccgggct gtccccgcac gctgccggct cggggatgcg gggggagcgc
     6601 cggaccggag cggagccccg ggcggctcgc tgctgccccc tagcggggga gggacgtaat
     6661 tacatccctg ggggctttgg gggggggctg tccctgatat ctataacaag aaaatatata
     6721 tataataagt tatcacgtaa gtagaacatg aaataacaat ataattatcg tatgagttaa
     6781 atcttaaaag tcacgtaaaa gataatcatg cgtcattttg actcacgcgg tcgttatagt
     6841 tcaaaatcag tgacacttac cgcattgaca agcacgcctc acgggagctc caagcggcga
     6901 ctgagatgtc ctaaatgcac agcgacggat tcgcgctatt tagaaagaga gagcaatatt
     6961 tcaagaatgc atgcgtcaat tttacgcaga ctatctttct agggttaatc tagctgcatc
     7021 aggatcatat cgtcgggtct tttttccggc tcagtcatcg cccaagctgg cgctatctgg
     7081 gcatcgggga ggaagaagcc cgtgcctttt cccgcgaggt tgaagcggca tggaaagagt
     7141 ttgccgagga tgactgctgc tgcattgacg ttgagcgaaa acgcacgttt accatgatga
     7201 ttcgggaagg tgtggccatg cacgccttta acggtgaact gttcgttcag gccacctggg
     7261 ataccagttc gtcgcggctt ttccggacac agttccggat ggtcagcccg aagcgcatca
     7321 gcaacccgaa caataccggc gacagccgga actgccgtgc cggtgtgcag attaatgaca
     7381 gcggtgcggc gctgggatat tacgtcagcg aggacgggta tcctggctgg atgccgcaga
     7441 aatggacatg gataccccgt gagttacccg gcgggcgcgc ttggcgtaat catggtcata
     7501 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     7561 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     7621 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     7681 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     7741 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     7801 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     7861 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     7921 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     7981 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     8041 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc atagctcacg
     8101 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     8161 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     8221 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     8281 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     8341 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     8401 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     8461 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     8521 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     8581 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     8641 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     8701 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     8761 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     8821 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     8881 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     8941 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     9001 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     9061 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     9121 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     9181 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     9241 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     9301 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     9361 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     9421 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     9481 gggaataagg gcgacacgga aatgttgaat actcat
//
