LOCUS       62056_12765        7842 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 7842)
  AUTHORS   .
  TITLE     Direct Submission
FEATURES             Location/Qualifiers
     source          1..7842
                     /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"
     promoter        1389..1600
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor 
                     EF-1-alpha"
     LTR             1613..1881
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2722..2751
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     CDS             3625..3678
                     /codon_start=1
                     /label=T2A
                     /note="2A peptide from Thosea asigna virus capsid protein"
                     /translation="EGRGSLLTCGDVEENPGP"
     CDS             3685..4281
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    4412..4544
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(5821..5837)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    complement(5845..5861)
                     /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(5869..5899)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    complement(5914..5935)
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(6223..6811)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(6985..7842)
                     /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 ataatcatgt gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaac aaaaactcaa
     1081 aatttcttct ataaagtaac aaaactttta tgagggacag ccccccccca aagcccccag
     1141 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     1201 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     1261 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     1321 cacctggggg gatacgggga aaaggcctcc acggccaagg atctgcgatc gctccggtgc
     1381 ccgtcagtgg gcagagcgca catcgcccac agtccccgag aagttggggg gaggggtcgg
     1441 caattgaacg ggtgcctaga gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta
     1501 ctggctccgc ctttttcccg agggtggggg agaaccgtat ataagtgcag tagtcgccgt
     1561 gaacgttctt tttcgcaacg ggtttgccgc cagaacacag ctgaagcttc gaggggctcg
     1621 catctctcct tcacgcgccc gccgccctac ctgaggccgc catccacgcc ggttgagtcg
     1681 cgttctgccg cctcccgcct gtggtgcctc ctgaactgcg tccgccgtct aggtaagttt
     1741 aaagctcagg tcgagaccgg gcctttgtcc ggcgctccct tggagcctac ctagactcag
     1801 ccggctctcc acgctttgcc tgaccctgct tgctcaactc tacgtctttg tttcgttttc
     1861 tgttctgcgc cgttacagat ccaagctgtg accggcgcct actctagagc caccatgctg
     1921 ctgctggtca cttctctgct gctgtgcgaa ctgccccacc ccgcctttct gctgattccc
     1981 caggtgcagc tggtgcaatc tggggctgag gtgaagaagc ctgggtcctc ggtgaaggtc
     2041 tcctgcaagg cttctggagg caccctcagc agctatgcta tcagctgggt gcgacaggcc
     2101 cctggacaag ggcctgagtg gatgggatgg atcaacccta atagtggtgg cacaaactat
     2161 gcacagaagt ttcagggcag ggtcaccatg accagggaca cgtccatcag cacagcctac
     2221 atggagctga gcaggctgag acctgacgac acggccgtgt attactgtgc gagtggctat
     2281 tgtagtagta ccagctgcta tgactactgg ggccagggca ccctggtcac cgtctcctca
     2341 ggcagcggcg gtggcggatc ccaggctgtg ctcactcagc cgtcttccct ctctgcatct
     2401 cctggagcat cagtcagtct cacctgcact ttacgcagtg gcatcaatgt tggtgcctac
     2461 aggatatact ggtaccagca gaagccaggg agtcctcccc agtttctcct gaggtacaaa
     2521 tcagactcag ataagcagca gggctctgga gtccccagcc gcttctctgg atccagagat
     2581 gcttcggcca atgcagggat tttactcatc tctgggctcc ggtctgagga tgaggctgac
     2641 tattactgtg cgatttggca cagcagcgct tgggtgttcg gcggagggac ccagctcacc
     2701 gttttaggtg ccgcaggcag cgaacagaaa ctgatttccg aggaagatct gttcgtcccc
     2761 gtgttcctgc ctgccaagcc aacaactacc cctgctccac gaccacctac tccagcacct
     2821 accatcgcaa gtcagcccct gtcactgcga cctgaggctt gccggccagc agctggagga
     2881 gcagtgcaca cccgaggcct ggacttcgca tgcgatatct acatttgggc accactggct
     2941 ggaacctgtg gggtcctgct gctgagcctg gtcatcaccc tgtattgtaa ccacagaaat
     3001 aggagcaaac gctcccgact gctgcattcc gactacatga acatgacacc tcggagacca
     3061 ggccccacta gaaagcatta ccagccatat gccccaccca gggatttcgc agcctatcgg
     3121 agccggttca gcgtcgtgaa aagggggcgc aagaaactgc tgtacatctt caagcagcct
     3181 tttatgcgcc cagtgcagac aactcaggag gaagacggat gctcttgtcg gttcccagag
     3241 gaggaggaag gaggctgcga gctgagagtg aagttcagcc ggagcgccga tgcaccagca
     3301 tatcagcagg gacagaatca gctgtacaac gagctgaatc tgggcaggcg cgaggaatat
     3361 gacgtgctgg ataagcgacg aggacgggac cccgaaatgg gaggaaaacc cagaaggaag
     3421 aaccctcagg aggggctgta taatgaactg cagaaagaca agatggctga ggcatacagc
     3481 gaaattggaa tgaaaggaga gcgccgacgg gggaagggac acgatgggct gtaccaggga
     3541 ctgtcaaccg ccactaaaga tacctacgac gcactgcaca tgcaggctct gcccccaaga
     3601 gaattcgaag gatccgcggc cgctgagggc agaggaagtc ttctaacatg cggtgacgtg
     3661 gaggagaatc ccggcccttc cgggatgacc gagtacaagc ccacggtgcg cctcgccacc
     3721 cgcgacgacg tccccagggc cgtacgcacc ctcgccgccg cgttcgccga ctaccccgcc
     3781 acgcgccaca ccgtcgatcc ggaccgccac atcgagcggg tcaccgagct gcaagaactc
     3841 ttcctcacgc gcgtcgggct cgacatcggc aaggtgtggg tcgcggacga cggcgccgcg
     3901 gtggcggtct ggaccacgcc ggagagcgtc gaagcggggg cggtgttcgc cgagatcggc
     3961 ccgcgcatgg ccgagttgag cggttcccgg ctggccgcgc agcaacagat ggaaggcctc
     4021 ctggcgccgc accggcccaa ggagcccgcg tggttcctgg ccaccgtcgg cgtctcgccc
     4081 gaccaccagg gcaagggtct gggcagcgcc gtcgtgctcc ccggagtgga ggcggccgag
     4141 cgcgccgggg tgcccgcctt cctggagacc tccgcgcccc gcaacctccc cttctacgag
     4201 cggctcggct tcaccgtcac cgccgacgtc gaggtgcccg aaggaccgcg cacctggtgc
     4261 atgacccgca agcccggtgc ctgaatctag gtcgacaatc aacctctgga ttacaaaatt
     4321 tgtgaaagat tgactggtat tcttaactat gttgctcctt ttacgctatg tggatacgct
     4381 gctttaatgc ctttgtatca tgcgttaact aaacttgttt attgcagctt ataatggtta
     4441 caaataaagc aatagcatca caaatttcac aaataaagca tttttttcac tgcattctag
     4501 ttgtggtttg tccaaactca tcaatgtatc ttatcatgtc tggaattgac tcaaatgatg
     4561 tcaattagtc tatcagaagc tcatctggtc tcccttccgg gggacaagac atccctgttt
     4621 aatatttaaa cagcagtgtt cccaaactgg gttcttatat cccttgctct ggtcaaccag
     4681 gttgcagggt ttcctgtcct cacaggaacg aagtccctaa agaaacagtg gcagccaggt
     4741 ttagccccgg aattgactgg attccttttt tagggcccat tggtatggct ttttccccgt
     4801 atccccccag gtgtctgcag gctcaaagag cagcgagaag cgttcagagg aaagcgatcc
     4861 cgtgccacct tccccgtgcc cgggctgtcc ccgcacgctg ccggctcggg gatgcggggg
     4921 gagcgccgga ccggagcgga gccccgggcg gctcgctgct gccccctagc gggggaggga
     4981 cgtaattaca tccctggggg ctttgggggg gggctgtccc tgatatctat aacaagaaaa
     5041 tatatatata ataagttatc acgtaagtag aacatgaaat aacaatataa ttatcgtatg
     5101 agttaaatct taaaagtcac gtaaaagata atcatgcgtc attttgactc acgcggtcgt
     5161 tatagttcaa aatcagtgac acttaccgca ttgacaagca cgcctcacgg gagctccaag
     5221 cggcgactga gatgtcctaa atgcacagcg acggattcgc gctatttaga aagagagagc
     5281 aatatttcaa gaatgcatgc gtcaatttta cgcagactat ctttctaggg ttaatctagc
     5341 tgcatcagga tcatatcgtc gggtcttttt tccggctcag tcatcgccca agctggcgct
     5401 atctgggcat cggggaggaa gaagcccgtg ccttttcccg cgaggttgaa gcggcatgga
     5461 aagagtttgc cgaggatgac tgctgctgca ttgacgttga gcgaaaacgc acgtttacca
     5521 tgatgattcg ggaaggtgtg gccatgcacg cctttaacgg tgaactgttc gttcaggcca
     5581 cctgggatac cagttcgtcg cggcttttcc ggacacagtt ccggatggtc agcccgaagc
     5641 gcatcagcaa cccgaacaat accggcgaca gccggaactg ccgtgccggt gtgcagatta
     5701 atgacagcgg tgcggcgctg ggatattacg tcagcgagga cgggtatcct ggctggatgc
     5761 cgcagaaatg gacatggata ccccgtgagt tacccggcgg gcgcgcttgg cgtaatcatg
     5821 gtcatagctg tttcctgtgt gaaattgtta tccgctcaca attccacaca acatacgagc
     5881 cggaagcata aagtgtaaag cctggggtgc ctaatgagtg agctaactca cattaattgc
     5941 gttgcgctca ctgcccgctt tccagtcggg aaacctgtcg tgccagctgc attaatgaat
     6001 cggccaacgc gcggggagag gcggtttgcg tattgggcgc tcttccgctt cctcgctcac
     6061 tgactcgctg cgctcggtcg ttcggctgcg gcgagcggta tcagctcact caaaggcggt
     6121 aatacggtta tccacagaat caggggataa cgcaggaaag aacatgtgag caaaaggcca
     6181 gcaaaaggcc aggaaccgta aaaaggccgc gttgctggcg tttttccata ggctccgccc
     6241 ccctgacgag catcacaaaa atcgacgctc aagtcagagg tggcgaaacc cgacaggact
     6301 ataaagatac caggcgtttc cccctggaag ctccctcgtg cgctctcctg ttccgaccct
     6361 gccgcttacc ggatacctgt ccgcctttct cccttcggga agcgtggcgc tttctcatag
     6421 ctcacgctgt aggtatctca gttcggtgta ggtcgttcgc tccaagctgg gctgtgtgca
     6481 cgaacccccc gttcagcccg accgctgcgc cttatccggt aactatcgtc ttgagtccaa
     6541 cccggtaaga cacgacttat cgccactggc agcagccact ggtaacagga ttagcagagc
     6601 gaggtatgta ggcggtgcta cagagttctt gaagtggtgg cctaactacg gctacactag
     6661 aaggacagta tttggtatct gcgctctgct gaagccagtt accttcggaa aaagagttgg
     6721 tagctcttga tccggcaaac aaaccaccgc tggtagcggt ggtttttttg tttgcaagca
     6781 gcagattacg cgcagaaaaa aaggatctca agaagatcct ttgatctttt ctacggggtc
     6841 tgacgctcag tggaacgaaa actcacgtta agggattttg gtcatgagat tatcaaaaag
     6901 gatcttcacc tagatccttt taaattaaaa atgaagtttt aaatcaatct aaagtatata
     6961 tgagtaaact tggtctgaca gttaccaatg cttaatcagt gaggcaccta tctcagcgat
     7021 ctgtctattt cgttcatcca tagttgcctg actccccgtc gtgtagataa ctacgatacg
     7081 ggagggctta ccatctggcc ccagtgctgc aatgataccg cgagacccac gctcaccggc
     7141 tccagattta tcagcaataa accagccagc cggaagggcc gagcgcagaa gtggtcctgc
     7201 aactttatcc gcctccatcc agtctattaa ttgttgccgg gaagctagag taagtagttc
     7261 gccagttaat agtttgcgca acgttgttgc cattgctaca ggcatcgtgg tgtcacgctc
     7321 gtcgtttggt atggcttcat tcagctccgg ttcccaacga tcaaggcgag ttacatgatc
     7381 ccccatgttg tgcaaaaaag cggttagctc cttcggtcct ccgatcgttg tcagaagtaa
     7441 gttggccgca gtgttatcac tcatggttat ggcagcactg cataattctc ttactgtcat
     7501 gccatccgta agatgctttt ctgtgactgg tgagtactca accaagtcat tctgagaata
     7561 gtgtatgcgg cgaccgagtt gctcttgccc ggcgtcaata cgggataata ccgcgccaca
     7621 tagcagaact ttaaaagtgc tcatcattgg aaaacgttct tcggggcgaa aactctcaag
     7681 gatcttaccg ctgttgagat ccagttcgat gtaacccact cgtgcaccca actgatcttc
     7741 agcatctttt actttcacca gcgtttctgg gtgagcaaaa acaggaaggc aaaatgccgc
     7801 aaaaaaggga ataagggcga cacggaaatg ttgaatactc at
//
