LOCUS       Exported                9743 bp ds-DNA     circular SYN 12-MAY-2021
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    pT3-EF1a-c-Met
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 9743)
  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 9743)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported May 12, 2021 from SnapGene Server 1.1.58
            http://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..9743
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     primer_bind     174..193
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     427..444
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    561..582
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        597..627
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    635..651
                     /label=lac operator
                     /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-beta-D-thiogalactopyranoside (IPTG)."
     primer_bind     640..662
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     659..675
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     659..675
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1101..1120)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     protein_bind    complement(1173..1206)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     promoter        1347..2525
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          1578..2516
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     2473..2493
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
     primer_bind     2542..2561
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2542..2560
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     protein_bind    2639..2663
                     /gene="mutant version of attB"
                     /label=attB1
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     protein_bind    complement(6855..6879)
                     /gene="mutant version of attB"
                     /label=attB2
                     /bound_moiety="BP Clonase(TM)"
                     /note="recombination site for the Gateway(R) BP reaction"
     CDS             6932..6973
                     /codon_start=1
                     /product="epitope tag from simian virus 5"
                     /label=V5 tag
                     /translation="GKPIPNPLLGLDST"
     primer_bind     complement(7011..7028)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    7017..7241
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    complement(7311..7344)
                     /label=loxP
                     /bound_moiety="Cre recombinase"
                     /note="Cre-mediated recombination occurs in the 8-bp core 
                     sequence (GCATACAT)."
     primer_bind     7400..7420
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     primer_bind     complement(7802..7819)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(7802..7818)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(7811..7833)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     complement(8027..8046)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     8146..8168
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     primer_bind     complement(8206..8224)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        8292..8396
                     /gene="bla"
                     /label=AmpR promoter
     CDS             8397..9257
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(8615..8634)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      9428..273
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 tgaacggggg gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga
       61 tacctacagc gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg
      121 tatccggtaa gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac
      181 gcctggtatc tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg
      241 tgatgctcgt caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg
      301 ttcctggcct tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct
      361 gtggataacc gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc
      421 gagcgcagcg agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc
      481 cccgcgcgtt ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg
      541 ggcagtgagc gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta
      601 cactttatgc ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca
      661 ggaaacagct atgaccatga ttacgccaag cttgcatgcc tgcaggtcga ctctagacct
      721 atacagttga agtcggaagt ttacatacac ttaagttgga gtcattaaaa ctcgtttttc
      781 aactactcca caaatttctt gttaacaaac aatagttttg gcaagtcagt taggacatct
      841 actttgtgca tgacacaagt catttttcca acaattgttt acagacagat tatttcactt
      901 ataattcact gtatcacaat tccagtgggt cagaagttta catacactaa gttgactgtg
      961 cctttaaaca gcttggaaaa ttccagaaaa tgatgtcatg gctttagaag cttctgatag
     1021 actaattgac atcatttgag tcaattggag gtgtacctgt ggatgtattt caaggggatc
     1081 cagacatgat aagatacatt gatgagtttg gacaaaccac aactagaatg cagcggccgc
     1141 tagctcgaga tcttaattaa actagtatcg atataacttc gtataatgta tgctatacga
     1201 agttatatgt acgggccaga tatacgcgtt gacattgatt attgactagg cttttgcaaa
     1261 aagctttgca aagatggata aagttttaaa cagagaggaa tctttgcagc taatggacct
     1321 tctaggtctt gaaaggagtg ggaattggct ccggtgcccg tcagtgggca gagcgcacat
     1381 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaaccggt gcctagagaa
     1441 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     1501 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     1561 ttgccgccag aacacaggta agtgccgtgt gtggttcccg cgggcctggc ctctttacgg
     1621 gttatggccc ttgcgtgcct tgaattactt ccacctggct gcagtacgtg attcttgatc
     1681 ccgagcttcg ggttggaagt gggtgggaga gttcgaggcc ttgcgcttaa ggagcccctt
     1741 cgcctcgtgc ttgagttgag gcctggcctg ggcgctgggg ccgccgcgtg cgaatctggt
     1801 ggcaccttcg cgcctgtctc gctgctttcg ataagtctct agccatttaa aatttttgat
     1861 gacctgctgc gacgcttttt ttctggcaag atagtcttgt aaatgcgggc caagatctgc
     1921 acactggtat ttcggttttt ggggccgcgg gcggcgacgg ggcccgtgcg tcccagcgca
     1981 catgttcggc gaggcggggc ctgcgagcgc ggccaccgag aatcggacgg gggtagtctc
     2041 aagctggccg gcctgctctg gtgcctggcc tcgcgccgcc gtgtatcgcc ccgccctggg
     2101 cggcaaggct ggcccggtcg gcaccagttg cgtgagcgga aagatggccg cttcccggcc
     2161 ctgctgcagg gagctcaaaa tggaggacgc ggcgctcggg agagcgggcg ggtgagtcac
     2221 ccacacaaag gaaaagggcc tttccgtcct cagccgtcgc ttcatgtgac tccacggagt
     2281 accgggcgcc gtccaggcac ctcgattagt tctcgagctt ttggagtacg tcgtctttag
     2341 gttgggggga ggggttttat gcgatggagt ttccccacac tgagtgggtg gagactgaag
     2401 ttaggccagc ttggcacttg atgtaattct ccttggaatt tgcccttttt gagtttggat
     2461 cttggttcat tctcaagcct cagacagtgg ttcaaagttt ttttcttcca tttcaggtgt
     2521 cgtgaggaat tagcttggta ctaatacgac tcactatagg gagacccaag ctggctaggt
     2581 aagcttggta ccgagctcgg atccactagt ccagtgtggt ggaattctgc agatatcaac
     2641 aagtttgtac aaaaaagcag gcttcaaaga taaacctctc ataatgaagg cccccgctgt
     2701 gcttgcacct ggcatcctcg tgctcctgtt taccttggtg cagaggagca atggggagtg
     2761 taaagaggca ctagcaaagt ccgagatgaa tgtgaatatg aagtatcagc ttcccaactt
     2821 caccgcggaa acacccatcc agaatgtcat tctacatgag catcacattt tccttggtgc
     2881 cactaactac atttatgttt taaatgagga agaccttcag aaggttgctg agtacaagac
     2941 tgggcctgtg ctggaacacc cagattgttt cccatgtcag gactgcagca gcaaagccaa
     3001 tttatcagga ggtgtttgga aagataacat caacatggct ctagttgtcg acacctacta
     3061 tgatgatcaa ctcattagct gtggcagcgt caacagaggg acctgccagc gacatgtctt
     3121 tccccacaat catactgctg acatacagtc ggaggttcac tgcatattct ccccacagat
     3181 agaagagccc agccagtgtc ctgactgtgt ggtgagcgcc ctgggagcca aagtcctttc
     3241 atctgtaaag gaccggttca tcaacttctt tgtaggcaat accataaatt cttcttattt
     3301 cccagatcat ccattgcatt cgatatcagt gagaaggcta aaggaaacga aagatggttt
     3361 tatgtttttg acggaccagt cctacattga tgttttacct gagttcagag attcttaccc
     3421 cattaagtat gtccatgcct ttgaaagcaa caattttatt tacttcttga cggtccaaag
     3481 ggaaactcta gatgctcaga cttttcacac aagaataatc aggttctgtt ccataaactc
     3541 tggattgcat tcctacatgg aaatgcctct ggagtgtatt ctcacagaaa agagaaaaaa
     3601 gagatccaca aagaaggaag tgtttaatat acttcaggct gcgtatgtca gcaagcctgg
     3661 ggcccagctt gctagacaaa taggagccag cctgaatgat gacattcttt tcggggtgtt
     3721 cgcacaaagc aagccagatt ctgccgaacc aatggatcga tctgccatgt gtgcattccc
     3781 tatcaaatat gtcaacgact tcttcaacaa gatcgtcaac aaaaacaatg tgagatgtct
     3841 ccagcatttt tacggaccca atcatgagca ctgctttaat aggacacttc tgagaaattc
     3901 atcaggctgt gaagcgcgcc gtgatgaata tcgaacagag tttaccacag ctttgcagcg
     3961 cgttgactta ttcatgggtc aattcagcga agtcctctta acatctatat ccaccttcat
     4021 taaaggagac ctcaccatag ctaatcttgg gacatcagag ggtcgcttca tgcaggttgt
     4081 ggtttctcga tcaggaccat caacccctca tgtgaatttt ctcctggact cccatccagt
     4141 gtctccagaa gtgattgtgg agcatacatt aaaccaaaat ggctacacac tggttatcac
     4201 tgggaagaag atcacgaaga tcccattgaa tggcttgggc tgcagacatt tccagtcctg
     4261 cagtcaatgc ctctctgccc caccctttgt tcagtgtggc tggtgccacg acaaatgtgt
     4321 gcgatcggag gaatgcctga gcgggacatg gactcaacag atctgtctgc ctgcaatcta
     4381 caaggttttc ccaaatagtg caccccttga aggagggaca aggctgacca tatgtggctg
     4441 ggactttgga tttcggagga ataataaatt tgatttaaag aaaactagag ttctccttgg
     4501 aaatgagagc tgcaccttga ctttaagtga gagcacgatg aatacattga aatgcacagt
     4561 tggtcctgcc atgaataagc atttcaatat gtccataatt atttcaaatg gccacgggac
     4621 aacacaatac agtacattct cctatgtgga tcctgtaata acaagtattt cgccgaaata
     4681 cggtcctatg gctggtggca ctttacttac tttaactgga aattacctaa acagtgggaa
     4741 ttctagacac atttcaattg gtggaaaaac atgtacttta aaaagtgtgt caaacagtat
     4801 tcttgaatgt tataccccag cccaaaccat ttcaactgag tttgctgtta aattgaaaat
     4861 tgacttagcc aaccgagaga caagcatctt cagttaccgt gaagatccca ttgtctatga
     4921 aattcatcca accaaatctt ttattagtgg tgggagcaca ataacaggtg ttgggaaaaa
     4981 cctgaattca gttagtgtcc cgagaatggt cataaatgtg catgaagcag gaaggaactt
     5041 tacagtggca tgtcaacatc gctctaattc agagataatc tgttgtacca ctccttccct
     5101 gcaacagctg aatctgcaac tccccctgaa aaccaaagcc tttttcatgt tagatgggat
     5161 cctttccaaa tactttgatc tcatttatgt acataatcct gtgtttaagc cttttgaaaa
     5221 gccagtgatg atctcaatgg gcaatgaaaa tgtactggaa attaagggaa atgatattga
     5281 ccctgaagca gttaaaggtg aagtgttaaa agttggaaat aagagctgtg agaatataca
     5341 cttacattct gaagccgttt tatgcacggt ccccaatgac ctgctgaaat tgaacagcga
     5401 gctaaatata gagtggaagc aagcaatttc ttcaaccgtc cttggaaaag taatagttca
     5461 accagatcag aatttcacag gattgattgc tggtgttgtc tcaatatcaa cagcactgtt
     5521 attactactt gggtttttcc tgtggctgaa aaagagaaag caaattaaag atctgggcag
     5581 tgaattagtt cgctacgatg caagagtaca cactcctcat ttggataggc ttgtaagtgc
     5641 ccgaagtgta agcccaacta cagaaatggt ttcaaatgaa tctgtagact accgagctac
     5701 ttttccagaa gatcagtttc ctaattcatc tcagaacggt tcatgccgac aagtgcagta
     5761 tcctctgaca gacatgtccc ccatcctaac tagtggggac tctgatatat ccagtccatt
     5821 actgcaaaat actgtccaca ttgacctcag tgctctaaat ccagagctgg tccaggcagt
     5881 gcagcatgta gtgattgggc ccagtagcct gattgtgcat ttcaatgaag tcataggaag
     5941 agggcatttt ggttgtgtat atcatgggac tttgttggac aatgatggca agaaaattca
     6001 ctgtgctgtg aaatccttga acagaatcac tgacatagga gaagtttccc aatttctgac
     6061 cgagggaatc atcatgaaag attttagtca tcccaatgtc ctctcgctcc tgggaatctg
     6121 cctgcgaagt gaagggtctc cgctggtggt cctaccatac atgaaacatg gagatcttcg
     6181 aaatttcatt cgaaatgaga ctcataatcc aactgtaaaa gatcttattg gctttggtct
     6241 tcaagtagcc aaaggcatga aatatcttgc aagcaaaaag tttgtccaca gagacttggc
     6301 tgcaagaaac tgtatgctgg atgaaaaatt cacagtcaag gttgctgatt ttggtcttgc
     6361 cagagacatg tatgataaag aatactatag tgtacacaac aaaacaggtg caaagctgcc
     6421 agtgaagtgg atggctttgg aaagtctgca aactcaaaag tttaccacca agtcagatgt
     6481 gtggtccttt ggcgtgctcc tctgggagct gatgacaaga ggagccccac cttatcctga
     6541 cgtaaacacc tttgatataa ctgtttactt gttgcaaggg agaagactcc tacaacccga
     6601 atactgccca gaccccttat atgaagtaat gctaaaatgc tggcacccta aagccgaaat
     6661 gcgcccatcc ttttctgaac tggtgtcccg gatatcagcg atcttctcta ctttcattgg
     6721 ggagcactat gtccatgtga acgctactta tgtgaacgta aaatgtgtcg ctccgtatcc
     6781 ttctctgttg tcatcagaag ataacgctga tgatgaggtg gacacacgac cagcctcctt
     6841 ctgggagaca tcagacccag ctttcttgta caaagtggtt gatatccagc acagtggcgg
     6901 ccgctcgagt ctagagggcc cgcggttcga aggtaagcct atccctaacc ctctcctcgg
     6961 tctcgattct acgcgtaccg gttagtaatg agtttaaacc cgctgatcag cctcgactgt
     7021 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
     7081 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
     7141 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
     7201 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
     7261 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa ataacttcgt
     7321 ataatgtatg ctatacgaag ttatatcgta tgcatgcttt gcatacttct gcctgctggg
     7381 gagcctgggg actttccaca ccctaactga cacacattcc acagaattcc ttgaaataca
     7441 tccacaggta cacctccaat tgactcaaat gatgtcaatt agtctatcag aagcttctaa
     7501 agccatgaca tcattttctg gaattttcca agctgtttaa aggcacagtc aacttagtgt
     7561 atgtaaactt ctgacccact ggaattgtga tacagtgaat tataagtgaa ataatctgtc
     7621 tgtaaacaat tgttggaaaa atgacttgtg tcatgcacaa agtagatgtc ctaactgact
     7681 tgccaaaact attgtttgtt aacaagaaat ttgtggagta gttgaaaaac gagttttaat
     7741 gactccaact taagtgtatg taaacttccg acttcaactg tagggtaccg agctcgaatt
     7801 cactggccgt cgttttacaa cgtcgtgact gggaaaaccc tggcgttacc caacttaatc
     7861 gccttgcagc acatccccct ttcgccagct ggcgtaatag cgaagaggcc cgcaccgatc
     7921 gcccttccca acagttgcgc agcctgaatg gcgaatggcg cctgatgcgg tattttctcc
     7981 ttacgcatct gtgcggtatt tcacaccgca tatggtgcac tctcagtaca atctgctctg
     8041 atgccgcata gttaagccag ccccgacacc cgccaacacc cgctgacgcg ccctgacggg
     8101 cttgtctgct cccggcatcc gcttacagac aagctgtgac cgtctccggg agctgcatgt
     8161 gtcagaggtt ttcaccgtca tcaccgaaac gcgcgagacg aaagggcctc gtgatacgcc
     8221 tatttttata ggttaatgtc atgataataa tggtttctta gacgtcaggt ggcacttttc
     8281 ggggaaatgt gcgcggaacc cctatttgtt tatttttcta aatacattca aatatgtatc
     8341 cgctcatgag acaataaccc tgataaatgc ttcaataata ttgaaaaagg aagagtatga
     8401 gtattcaaca tttccgtgtc gcccttattc ccttttttgc ggcattttgc cttcctgttt
     8461 ttgctcaccc agaaacgctg gtgaaagtaa aagatgctga agatcagttg ggtgcacgag
     8521 tgggttacat cgaactggat ctcaacagcg gtaagatcct tgagagtttt cgccccgaag
     8581 aacgttttcc aatgatgagc acttttaaag ttctgctatg tggcgcggta ttatcccgta
     8641 ttgacgccgg gcaagagcaa ctcggtcgcc gcatacacta ttctcagaat gacttggttg
     8701 agtactcacc agtcacagaa aagcatctta cggatggcat gacagtaaga gaattatgca
     8761 gtgctgccat aaccatgagt gataacactg cggccaactt acttctgaca acgatcggag
     8821 gaccgaagga gctaaccgct tttttgcaca acatggggga tcatgtaact cgccttgatc
     8881 gttgggaacc ggagctgaat gaagccatac caaacgacga gcgtgacacc acgatgcctg
     8941 tagcaatggc aacaacgttg cgcaaactat taactggcga actacttact ctagcttccc
     9001 ggcaacaatt aatagactgg atggaggcgg ataaagttgc aggaccactt ctgcgctcgg
     9061 cccttccggc tggctggttt attgctgata aatctggagc cggtgagcgt gggtctcgcg
     9121 gtatcattgc agcactgggg ccagatggta agccctcccg tatcgtagtt atctacacga
     9181 cggggagtca ggcaactatg gatgaacgaa atagacagat cgctgagata ggtgcctcac
     9241 tgattaagca ttggtaactg tcagaccaag tttactcata tatactttag attgatttaa
     9301 aacttcattt ttaatttaaa aggatctagg tgaagatcct ttttgataat ctcatgacca
     9361 aaatccctta acgtgagttt tcgttccact gagcgtcaga ccccgtagaa aagatcaaag
     9421 gatcttcttg agatcctttt tttctgcgcg taatctgctg cttgcaaaca aaaaaaccac
     9481 cgctaccagc ggtggtttgt ttgccggatc aagagctacc aactcttttt ccgaaggtaa
     9541 ctggcttcag cagagcgcag ataccaaata ctgttcttct agtgtagccg tagttaggcc
     9601 accacttcaa gaactctgta gcaccgccta catacctcgc tctgctaatc ctgttaccag
     9661 tggctgctgc cagtggcgat aagtcgtgtc ttaccgggtt ggactcaaga cgatagttac
     9721 cggataaggc gcagcggtcg ggc
//
