LOCUS       V031531                 8045 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V031531
VERSION     V031531
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     primer_bind     141..157
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 fwd"
     enhancer        318..697
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        698..901
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     CDS             945..968
                     /codon_start=1
                     /product="FLAG® epitope tag, followed by an enterokinase
                     cleavage site"
                     /transl_table=1
                     /translation="DYKDDDDK"
                     /label="FLAG"
     CDS             987..2774
                     /label="TCF7L2(NM_030756)"
                     /note="TCF7L2(NM_030756)"
                     /gene="TCF7L2"
     polyA_signal    2804..3426
                     /note="human growth hormone polyadenylation signal"
                     /label="hGH poly(A) signal"
     promoter        3468..3765
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3819..4613
                     /codon_start=1
                     /gene="aph(3')-II (or nptII)"
                     /note="confers resistance to neomycin, kanamycin, and G418
                     (Geneticin®)"
                     /product="aminoglycoside phosphotransferase from Tn5"
                     /transl_table=1
                     /translation="MIEQDGLHAGSPAAWVERLFGYDWAQQTIGCSDAAVFRLSAQGRP
                     VLFVKTDLSGALNELQDEAARLSWLATTGVPCAAVLDVVTEAGRDWLLLGEVPGQDLLS
                     SHLAPAEKVSIMADAMRRLHTLDPATCPFDHQAKHRIERARTRMEAGLVDQDDLDEEHQ
                     GLAPAELFARLKARMPDGEDLVVTHGDACLPNIMVENGRFSGFIDCGRLGVADRYQDIA
                     LATRDIAEELGGEWADRFLVLYGIAAPDSQRIAFYRLLDEFF*"
                     /label="NeoR/KanR"
     polyA_signal    5265..5399
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     promoter        complement(5405..5423)
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     primer_bind     complement(5437..5453)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    5461..5477
                     /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(5485..5515)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    5530..5551
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(5839..6427)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(6598..7458)
                     /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"
     promoter        complement(7459..7563)
                     /gene="bla"
                     /label="AmpR promoter"
     rep_origin      7590..8018
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
ORIGIN
        1 ccattcgcca ttcaggctgc gcaactgttg ggaagggcga tcggtgcggg cctcttcgct
       61 attacgccag ctggcgaaag ggggatgtgc tgcaaggcga ttaagttggg taacgccagg
      121 gttttcccag tcacgacgtt gtaaaacgac ggccagtgcc aagctgatct atacattgaa
      181 tcaatattgg caattagcca tattagtcat tggttatata gcataaatca atattggcta
      241 ttggccattg catacgttgt atctatatca taatatgtac atttatattg gctcatgtcc
      301 aatatgaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
      361 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
      421 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
      481 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
      541 ccacttggca gtacatcaag tgtatcatat gccaagtccg ccccctattg acgtcaatga
      601 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttacgggact ttcctacttg
      661 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacac
      721 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
      781 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaataaccc
      841 cgccccgttg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
      901 tcgtttagtg aaccgtcaga attaattcag atcgatctac catggactac aaagacgatg
      961 acgacaagct tgcggccgcg aattctatgc cgcagctgaa cggcggtgga ggggatgacc
     1021 taggcgccaa cgacgaactg atttccttca aagacgaggg cgaacaggag gagaagagct
     1081 ccgaaaactc ctcggcagag agggatttag ctgatgtcaa atcgtctcta gtcaatgaat
     1141 cagaaacgaa tcaaaacagc tcctccgatt ccgaggcgga aagacggcct ccgcctcgct
     1201 ccgaaagttt ccgagacaaa tcccgggaaa gtttggaaga agcggccaag aggcaagatg
     1261 gagggctctt taaggggcca ccgtatcccg gctacccctt catcatgatc cccgacctga
     1321 cgagccccta cctccccaac ggatcgctct cgcccaccgc ccgaacctat ctccagatga
     1381 aatggccact gcttgatgtc caggcaggga gcctccagag tagacaagcc ctcaaggatg
     1441 cccggtcccc atcaccggca cacattgtct ctaacaaagt gccagtggtg cagcaccctc
     1501 accatgtcca ccccctcacg cctcttatca cgtacagcaa tgaacacttc acgccgggaa
     1561 acccacctcc acacttacca gccgacgtag accccaaaac aggaatccca cggcctccgc
     1621 accctccaga tatatccccg tattacccac tatcgcctgg caccgtagga caaatccccc
     1681 atccgctagg atggttagta ccacagcaag gtcaaccagt gtacccaatc acgacaggag
     1741 gattcagaca cccctacccc acagctctga ccgtcaatgc ttccatgtcc aggttccctc
     1801 cccatatggt cccaccacat catacgctac acacgacggg cattccgcat ccggccatag
     1861 tcacaccaac agtcaaacag gaatcgtccc agagtgatgt cggctcactc catagttcaa
     1921 agcatcagga ctccaaaaag gaagaagaaa agaagaagcc ccacataaag aaacctctta
     1981 atgcattcat gttgtatatg aaggaaatga gagcaaaggt cgtagctgag tgcacgttga
     2041 aagaaagcgc ggccatcaac cagatccttg ggcggaggtg gcatgcactg tccagagaag
     2101 agcaagcgaa atactacgag ctggcccgga aggagcgaca gcttcatatg caactgtacc
     2161 ccggctggtc cgcgcgggat aactatggaa agaagaagaa gaggaaaagg gacaagcagc
     2221 cgggagagac caatgaacac agcgaatgtt tcctaaatcc ttgcctttca cttcctccga
     2281 ttacagacct gagcgctcct aagaaatgcc gagcgcgctt tggccttgat caacagaata
     2341 actggtgcgg cccttgcagg agaaaaaaaa agtgcgttcg ctacatacaa ggtgaaggca
     2401 gctgcctcag cccaccctct tcagatggaa gcttactaga ttcgcctccc ccctccccga
     2461 acctgctagg ctcccctccc cgagacgcca agtcacagac tgagcagacc cagcctctgt
     2521 cgctgtccct gaagcccgac cccctggccc acctgtccat gatgcctccg ccacccgccc
     2581 tcctgctcgc tgaggccacc cacaaggcct ccgccctctg tcccaacggg gccctggacc
     2641 tgcccccagc cgctttgcag cctgccgccc cctcctcatc aattgcacag ccgtcgactt
     2701 cttccttaca ttcccacagc tccctggccg ggacccagcc ccagccgctg tcgctcgtca
     2761 ccaagtcttt agaatagggt accagtcgac tctagaggat cccgggtggc atccctgtga
     2821 cccctcccca gtgcctctcc tggccctgga agttgccact ccagtgccca ccagccttgt
     2881 cctaataaaa ttaagttgca tcattttgtc tgactaggtg tccttctata atattatggg
     2941 gtggaggggg gtggtatgga gcaaggggca agttgggaag acaacctgta gggcctgcgg
     3001 ggtctattgg gaaccaagct ggagtgcagt ggcacaatct tggctcactg caatctccgc
     3061 ctcctgggtt caagcgattc tcctgcctca gcctcccgag ttgttgggat tccaggcatg
     3121 catgaccagg ctcagctaat ttttgttttt ttggtagaga cggggtttca ccatattggc
     3181 caggctggtc tccaactcct aatctcaggt gatctaccca ccttggcctc ccaaattgct
     3241 gggattacag gcgtgaacca ctgctccctt ccctgtcctt ctgattttaa aataactata
     3301 ccagcaggag gacgtccaga cacagcatag gctacctggc catgcccaac cggtgggaca
     3361 tttgagttgc ttgcttggca ctgtcctctc atgcgttggg tccactcagt agatgcctgt
     3421 tgaattgggt acgcggccag cttggctgtg gaatgtgtgt cagttagggt gtggaaagtc
     3481 cccaggctcc ccagcaggca gaagtatgca aagcatgcat ctcaattagt cagcaaccag
     3541 gtgtggaaag tccccaggct ccccagcagg cagaagtatg caaagcatgc atctcaatta
     3601 gtcagcaacc atagtcccgc ccctaactcc gcccatcccg cccctaactc cgcccagttc
     3661 cgcccattct ccgccccatg gctgactaat tttttttatt tatgcagagg ccgaggccgc
     3721 ctcggcctct gagctattcc agaagtagtg aggaggcttt tttggaggaa ttgatcagct
     3781 tgggatctga tcaagagaca ggatgaggat cgtttcgcat gattgaacaa gatggattgc
     3841 acgcaggttc tccggccgct tgggtggaga ggctattcgg ctatgactgg gcacaacaga
     3901 caatcggctg ctctgatgcc gccgtgttcc ggctgtcagc gcaggggcgc ccggttcttt
     3961 ttgtcaagac cgacctgtcc ggtgccctga atgaactgca ggacgaggca gcgcggctat
     4021 cgtggctggc cacgacgggc gttccttgcg cagctgtgct cgacgttgtc actgaagcgg
     4081 gaagggactg gctgctattg ggcgaagtgc cggggcagga tctcctgtca tctcaccttg
     4141 ctcctgccga gaaagtatcc atcatggctg atgcaatgcg gcggctgcat acgcttgatc
     4201 cggctacctg cccattcgac caccaagcga aacatcgcat cgagcgagca cgtactcgga
     4261 tggaagccgg tcttgtcgat caggatgatc tggacgaaga gcatcagggg ctcgcgccag
     4321 ccgaactgtt cgccaggctc aaggcgcgca tgcccgacgg cgaggatctc gtcgtgaccc
     4381 atggcgatgc ctgcttgccg aatatcatgg tggaaaatgg ccgcttttct ggattcatcg
     4441 actgtggccg gctgggtgtg gcggaccgct atcaggacat agcgttggct acccgtgata
     4501 ttgctgaaga gcttggcggc gaatgggctg accgcttcct cgtgctttac ggtatcgccg
     4561 ctcccgattc gcagcgcatc gccttctatc gccttcttga cgagttcttc tgagcgggac
     4621 tctggggttc gaaatgaccg accaagcgac gcccaacctg ccatcacgag atttcgattc
     4681 caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg ccggctggat
     4741 gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccgggc tcgatcccct
     4801 cgcgagttgg ttcagctgct gcctgaggct ggacgacctc gcggagttct accggcagtg
     4861 caaatccgtc ggcatccagg aaaccagcag cggctatccg cgcatccatg cccccgaact
     4921 gcaggagtgg ggaggcacga tggccgcttt ggtcgacccg gacgggacgc tcctgcgcct
     4981 gatacagaac gaattgcttg caggcatctc atgagtgtgt cttcccgttt tccgcctgag
     5041 gtcactgcgt ggatggagcg ctggcgcctg ctgcgcgacg gcgagctgct caccacccac
     5101 tcgccaagct ggaaccgtaa aaaggccgcg ttgctggcgt ttttccatag gctccgccga
     5161 tcataatcag ccataccaca tttgtagagg ttttacttgc tttaaaaaac ctcccacacc
     5221 tccccctgaa cctgaaacat aaaatgaatg caattgttgt tgttaacttg tttattgcag
     5281 cttataatgg ttacaaataa agcaatagca tcacaaattt cacaaataaa gcattttttt
     5341 cactgcattc tagttgtggt ttgtccaaac tcatcaatgt atcttatcat gtctggatca
     5401 attccctata gtgagtcgta ttaaattcgt aatcatgtca tagctgtttc ctgtgtgaaa
     5461 ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt gtaaagcctg
     5521 gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc ccgctttcca
     5581 gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg ggagaggcgg
     5641 tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct cggtcgttcg
     5701 gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca cagaatcagg
     5761 ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga accgtaaaaa
     5821 ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc acaaaaatcg
     5881 acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg cgtttccccc
     5941 tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat acctgtccgc
     6001 ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt atctcagttc
     6061 ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc agcccgaccg
     6121 ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg acttatcgcc
     6181 actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg gtgctacaga
     6241 gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg gtatctgcgc
     6301 tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg gcaaacaaac
     6361 caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca gaaaaaaagg
     6421 atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga acgaaaactc
     6481 acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga tccttttaaa
     6541 ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt ctgacagtta
     6601 ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt catccatagt
     6661 tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat ctggccccag
     6721 tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag caataaacca
     6781 gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct ccatccagtc
     6841 tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt tgcgcaacgt
     6901 tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg cttcattcag
     6961 ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca aaaaagcggt
     7021 tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt tatcactcat
     7081 ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat gcttttctgt
     7141 gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac cgagttgctc
     7201 ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa aagtgctcat
     7261 cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt tgagatccag
     7321 ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt tcaccagcgt
     7381 ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa gggcgacacg
     7441 gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt atcagggtta
     7501 ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa taggggttcc
     7561 gcgcacattt ccccgaaaag tgccacctga cgcgccctgt agcggcgcat taagcgcggc
     7621 gggtgtggtg gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc
     7681 tttcgctttc ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa
     7741 tcgggggctc cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact
     7801 tgattagggt gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt
     7861 gacgttggag tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa
     7921 ccctatctcg gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt
     7981 aaaaaatgag ctgatttaac aaaaatttaa cgcgaatttt aacaaaatat taacgcttac
     8041 aattt
//
