LOCUS       40924_42269        7681 bp DNA     circular SYN 26-MAY-2021
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7681)
  AUTHORS   Tward AD, Jones KD, Yant S, Cheung ST, Fan ST, Chen X, Kay MA, Wang 
            R, Bishop JM
  TITLE     Distinct pathways of genomic progression to benign and malignant 
            tumors of the liver.
  JOURNAL   Proc Natl Acad Sci U S A. 2007 Sep 11;104(37):14771-6. Epub 2007 Sep
            4.
  PUBMED    17785413
REFERENCE   2  (bases 1 to 7681)
  TITLE     Direct Submission
REFERENCE   3  (bases 1 to 7681)
  AUTHORS   .
  TITLE     Direct Submission
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Proc Natl
            Acad Sci U S A."; date: "2007-09-11"; volume: "104(37)"; pages: 
            "14771-6. Epub 2007 Sep 4"
COMMENT     SGRef: number: 2; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..7681
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        90..1268
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation
                     factor EF-1-alpha"
     promoter        1285..1303
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    1382..1406
                     /label=attB1
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             1417..1446
                     /codon_start=1
                     /label=Myc
                     /note="Myc (human c-Myc proto-oncogene) epitope tag"
                     /translation="EQKLISEEDL"
     protein_bind    complement(3536..3560)
                     /label=attB2
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             3613..3654
                     /codon_start=1
                     /label=V5 tag
                     /note="epitope tag from simian virus 5"
                     /translation="GKPIPNPLLGLDST"
     polyA_signal    3698..3922
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    3992..4025
                     /label=loxP
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (ATGTATGC) (Shaw et al., 2021)."
     primer_bind     4081..4101
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(4483..4499)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4708..4727)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable
                     marker"
     primer_bind     4827..4849
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(4887..4905)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward
                     primer"
     promoter        4973..5077
                     /label=AmpR promoter
     CDS             5078..5935
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      6109..6697
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     6851..6868
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    6985..7006
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        7021..7051
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    7059..7075
                     /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)."
     primer_bind     7083..7099
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7525..7544)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(7597..7630)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
ORIGIN
        1 aaaaagcttt gcaaagatgg ataaagtttt aaacagagag gaatctttgc agctaatgga
       61 ccttctaggt cttgaaagga gtgggaattg gctccggtgc ccgtcagtgg gcagagcgca
      121 catcgcccac agtccccgag aagttggggg gaggggtcgg caattgaacc ggtgcctaga
      181 gaaggtggcg cggggtaaac tgggaaagtg atgtcgtgta ctggctccgc ctttttcccg
      241 agggtggggg agaaccgtat ataagtgcag tagtcgccgt gaacgttctt tttcgcaacg
      301 ggtttgccgc cagaacacag gtaagtgccg tgtgtggttc ccgcgggcct ggcctcttta
      361 cgggttatgg cccttgcgtg ccttgaatta cttccacctg gctgcagtac gtgattcttg
      421 atcccgagct tcgggttgga agtgggtggg agagttcgag gccttgcgct taaggagccc
      481 cttcgcctcg tgcttgagtt gaggcctggc ctgggcgctg gggccgccgc gtgcgaatct
      541 ggtggcacct tcgcgcctgt ctcgctgctt tcgataagtc tctagccatt taaaattttt
      601 gatgacctgc tgcgacgctt tttttctggc aagatagtct tgtaaatgcg ggccaagatc
      661 tgcacactgg tatttcggtt tttggggccg cgggcggcga cggggcccgt gcgtcccagc
      721 gcacatgttc ggcgaggcgg ggcctgcgag cgcggccacc gagaatcgga cgggggtagt
      781 ctcaagctgg ccggcctgct ctggtgcctg gcctcgcgcc gccgtgtatc gccccgccct
      841 gggcggcaag gctggcccgg tcggcaccag ttgcgtgagc ggaaagatgg ccgcttcccg
      901 gccctgctgc agggagctca aaatggagga cgcggcgctc gggagagcgg gcgggtgagt
      961 cacccacaca aaggaaaagg gcctttccgt cctcagccgt cgcttcatgt gactccacgg
     1021 agtaccgggc gccgtccagg cacctcgatt agttctcgag cttttggagt acgtcgtctt
     1081 taggttgggg ggaggggttt tatgcgatgg agtttcccca cactgagtgg gtggagactg
     1141 aagttaggcc agcttggcac ttgatgtaat tctccttgga atttgccctt tttgagtttg
     1201 gatcttggtt cattctcaag cctcagacag tggttcaaag tttttttctt ccatttcagg
     1261 tgtcgtgagg aattagcttg gtactaatac gactcactat agggagaccc aagctggcta
     1321 ggtaagcttg gtaccgagct cggatccact agtccagtgt ggtggaattc tgcagatatc
     1381 aacaagtttg tacaaaaaag caggcttcgt accatggagc agaagctgat ctccgaggag
     1441 gacctgaagg atcctcgagc tcagagggta cgagctgcta tgttccctga gacattagat
     1501 gagggcatgc agatcccatc tacacagttt gatgctgctc atcccactaa tgtccagcgt
     1561 ttggctgaac catcacagat gctgaaacat gcagttgtaa acttgattaa ctatcaagat
     1621 gatgcagaac ttgccacacg tgcaatccct gaactgacaa aactgctaaa tgacgaggac
     1681 caggtggtgg ttaataaggc tgcagttatg gtccatcagc tttctaaaaa ggaagcttcc
     1741 agacacgcta tcatgcgttc tcctcagatg gtgtctgcta ttgtacgtac catgcagaat
     1801 acaaatgatg tagaaacagc tcgttgtacc gctgggacct tgcataacct ttcccatcat
     1861 cgtgagggct tactggccat ctttaagtct ggaggcattc ctgccctggt gaaaatgctt
     1921 ggttcaccag tggattctgt gttgttttat gccattacaa ctctccacaa ccttttatta
     1981 catcaagaag gagctaaaat ggcagtgcgt ttagctggtg ggctgcagaa aatggttgcc
     2041 ttgctcaaca aaacaaatgt taaattcttg gctattacga cagactgcct tcaaatttta
     2101 gcttatggca accaagaaag caagctcatc atactggcta gtggtggacc ccaagcttta
     2161 gtaaatataa tgaggaccta tacttacgaa aaactactgt ggaccacaag cagagtgctg
     2221 aaggtgctat ctgtctgctc tagtaataag ccggctattg tagaagctgg tggaatgcaa
     2281 gctttaggac ttcacctgac agatccaagt caacgtcttg ttcagaactg tctttggact
     2341 ctcaggaatc tttcagatgc tgcaactaaa caggaaggga tggaaggtct ccttgggact
     2401 cttgttcagc ttctgggttc agatgatata aatgtggtca cctgtgcagc tggaattctt
     2461 tctaacctca cttgcaataa ttataagaac aagatgatgg tctgccaagt gggtggtata
     2521 gaggctcttg tgcgtactgt ccttcgggct ggtgacaggg aagacatcac tgagcctgcc
     2581 atctgtgctc ttcgtcatct gaccagccga caccaagaag cagagatggc ccagaatgca
     2641 gttcgccttc actatggact accagttgtg gttaagctct tacacccacc atcccactgg
     2701 cctctgataa aggctactgt tggattgatt cgaaatcttg ccctttgtcc cgcaaatcat
     2761 gcacctttgc gtgagcaggg tgccattcca cgactagttc agttgcttgt tcgtgcacat
     2821 caggataccc agcgccgtac gtccatgggt gggacacagc agcaatttgt ggagggggtc
     2881 cgcatggaag aaatagttga aggttgtacc ggagcccttc acatcctagc tcgggatgtt
     2941 cacaaccgaa ttgttatcag aggactaaat accattccat tgtttgtgca gctgctttat
     3001 tctcccattg aaaacatcca aagagtagct gcaggggtcc tctgtgaact tgctcaggac
     3061 aaggaagctg cagaagctat tgaagctgag ggagccacag ctcctctgac agagttactt
     3121 cactctagga atgaaggtgt ggcgacatat gcagctgctg ttttgttccg aatgtctgag
     3181 gacaagccac aagattacaa gaaacggctt tcagttgagc tgaccagctc tctcttcaga
     3241 acagagccaa tggcttggaa tgagactgct gatcttggac ttgatattgg tgcccaggga
     3301 gaaccccttg gatatcgcca ggatgatcct agctatcgtt cttttcactc tggtggatat
     3361 ggccaggatg ccttgggtat ggaccccatg atggaacatg agatgggtgg ccaccaccct
     3421 ggtgctgact atccagttga tgggctgcca gatctggggc atgcccagga cctcatggat
     3481 gggctgcctc caggtgacag caatcagctg gcctggtttg atactgacct gtaagaccca
     3541 gctttcttgt acaaagtggt tgatatccag cacagtggcg gccgctcgag tctagagggc
     3601 ccgcggttcg aaggtaagcc tatccctaac cctctcctcg gtctcgattc tacgcgtacc
     3661 ggttagtaat gagtttaaac ccgctgatca gcctcgactg tgccttctag ttgccagcca
     3721 tctgttgttt gcccctcccc cgtgccttcc ttgaccctgg aaggtgccac tcccactgtc
     3781 ctttcctaat aaaatgagga aattgcatcg cattgtctga gtaggtgtca ttctattctg
     3841 gggggtgggg tggggcagga cagcaagggg gaggattggg aagacaatag caggcatgct
     3901 ggggatgcgg tgggctctat ggcttctgag gcggaaagaa ccagctgggg ctctaggggg
     3961 tatccccacg cgccctgtag cggcgcatta aataacttcg tataatgtat gctatacgaa
     4021 gttatatcgt atgcatgctt tgcatacttc tgcctgctgg ggagcctggg gactttccac
     4081 accctaactg acacacattc cacagaattc cttgaaatac atccacaggt acacctccaa
     4141 ttgactcaaa tgatgtcaat tagtctatca gaagcttcta aagccatgac atcattttct
     4201 ggaattttcc aagctgttta aaggcacagt caacttagtg tatgtaaact tctgacccac
     4261 tggaattgtg atacagtgaa ttataagtga aataatctgt ctgtaaacaa ttgttggaaa
     4321 aatgacttgt gtcatgcaca aagtagatgt cctaactgac ttgccaaaac tattgtttgt
     4381 taacaagaaa tttgtggagt agttgaaaaa cgagttttaa tgactccaac ttaagtgtat
     4441 gtaaacttcc gacttcaact gtagggtacc gagctcgaat tcactggccg tcgttttaca
     4501 acgtcgtgac tgggaaaacc ctggcgttac ccaacttaat cgccttgcag cacatccccc
     4561 tttcgccagc tggcgtaata gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg
     4621 cagcctgaat ggcgaatggc gcctgatgcg gtattttctc cttacgcatc tgtgcggtat
     4681 ttcacaccgc atatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     4741 gccccgacac ccgccaacac ccgctgacgc gccctgacgg gcttgtctgc tcccggcatc
     4801 cgcttacaga caagctgtga ccgtctccgg gagctgcatg tgtcagaggt tttcaccgtc
     4861 atcaccgaaa cgcgcgagac gaaagggcct cgtgatacgc ctatttttat aggttaatgt
     4921 catgataata atggtttctt agacgtcagg tggcactttt cggggaaatg tgcgcggaac
     4981 ccctatttgt ttatttttct aaatacattc aaatatgtat ccgctcatga gacaataacc
     5041 ctgataaatg cttcaataat attgaaaaag gaagagtatg agtattcaac atttccgtgt
     5101 cgcccttatt cccttttttg cggcattttg ccttcctgtt tttgctcacc cagaaacgct
     5161 ggtgaaagta aaagatgctg aagatcagtt gggtgcacga gtgggttaca tcgaactgga
     5221 tctcaacagc ggtaagatcc ttgagagttt tcgccccgaa gaacgttttc caatgatgag
     5281 cacttttaaa gttctgctat gtggcgcggt attatcccgt attgacgccg ggcaagagca
     5341 actcggtcgc cgcatacact attctcagaa tgacttggtt gagtactcac cagtcacaga
     5401 aaagcatctt acggatggca tgacagtaag agaattatgc agtgctgcca taaccatgag
     5461 tgataacact gcggccaact tacttctgac aacgatcgga ggaccgaagg agctaaccgc
     5521 ttttttgcac aacatggggg atcatgtaac tcgccttgat cgttgggaac cggagctgaa
     5581 tgaagccata ccaaacgacg agcgtgacac cacgatgcct gtagcaatgg caacaacgtt
     5641 gcgcaaacta ttaactggcg aactacttac tctagcttcc cggcaacaat taatagactg
     5701 gatggaggcg gataaagttg caggaccact tctgcgctcg gcccttccgg ctggctggtt
     5761 tattgctgat aaatctggag ccggtgagcg tgggtctcgc ggtatcattg cagcactggg
     5821 gccagatggt aagccctccc gtatcgtagt tatctacacg acggggagtc aggcaactat
     5881 ggatgaacga aatagacaga tcgctgagat aggtgcctca ctgattaagc attggtaact
     5941 gtcagaccaa gtttactcat atatacttta gattgattta aaacttcatt tttaatttaa
     6001 aaggatctag gtgaagatcc tttttgataa tctcatgacc aaaatccctt aacgtgagtt
     6061 ttcgttccac tgagcgtcag accccgtaga aaagatcaaa ggatcttctt gagatccttt
     6121 ttttctgcgc gtaatctgct gcttgcaaac aaaaaaacca ccgctaccag cggtggtttg
     6181 tttgccggat caagagctac caactctttt tccgaaggta actggcttca gcagagcgca
     6241 gataccaaat actgttcttc tagtgtagcc gtagttaggc caccacttca agaactctgt
     6301 agcaccgcct acatacctcg ctctgctaat cctgttacca gtggctgctg ccagtggcga
     6361 taagtcgtgt cttaccgggt tggactcaag acgatagtta ccggataagg cgcagcggtc
     6421 gggctgaacg gggggttcgt gcacacagcc cagcttggag cgaacgacct acaccgaact
     6481 gagataccta cagcgtgagc tatgagaaag cgccacgctt cccgaaggga gaaaggcgga
     6541 caggtatccg gtaagcggca gggtcggaac aggagagcgc acgagggagc ttccaggggg
     6601 aaacgcctgg tatctttata gtcctgtcgg gtttcgccac ctctgacttg agcgtcgatt
     6661 tttgtgatgc tcgtcagggg ggcggagcct atggaaaaac gccagcaacg cggccttttt
     6721 acggttcctg gccttttgct ggccttttgc tcacatgttc tttcctgcgt tatcccctga
     6781 ttctgtggat aaccgtatta ccgcctttga gtgagctgat accgctcgcc gcagccgaac
     6841 gaccgagcgc agcgagtcag tgagcgagga agcggaagag cgcccaatac gcaaaccgcc
     6901 tctccccgcg cgttggccga ttcattaatg cagctggcac gacaggtttc ccgactggaa
     6961 agcgggcagt gagcgcaacg caattaatgt gagttagctc actcattagg caccccaggc
     7021 tttacacttt atgcttccgg ctcgtatgtt gtgtggaatt gtgagcggat aacaatttca
     7081 cacaggaaac agctatgacc atgattacgc caagcttgca tgcctgcagg tcgactctag
     7141 acctatacag ttgaagtcgg aagtttacat acacttaagt tggagtcatt aaaactcgtt
     7201 tttcaactac tccacaaatt tcttgttaac aaacaatagt tttggcaagt cagttaggac
     7261 atctactttg tgcatgacac aagtcatttt tccaacaatt gtttacagac agattatttc
     7321 acttataatt cactgtatca caattccagt gggtcagaag tttacataca ctaagttgac
     7381 tgtgccttta aacagcttgg aaaattccag aaaatgatgt catggcttta gaagcttctg
     7441 atagactaat tgacatcatt tgagtcaatt ggaggtgtac ctgtggatgt atttcaaggg
     7501 gatccagaca tgataagata cattgatgag tttggacaaa ccacaactag aatgcagcgg
     7561 ccgctagctc gagatcttaa ttaaactagt atcgatataa cttcgtataa tgtatgctat
     7621 acgaagttat atgtacgggc cagatatacg cgttgacatt gattattgac taggcttttg
     7681 c
//
