LOCUS       V043473                 6897 bp    DNA     circular SYN 01-JAN-1980
DEFINITION  synthetic circular DNA.
ACCESSION   V043473
VERSION     V043473
KEYWORDS    .
SOURCE      .
  ORGANISM  .
            .
FEATURES             Location/Qualifiers
     enhancer        50..429
                     /note="human cytomegalovirus immediate early enhancer"
                     /label="CMV enhancer"
     promoter        430..633
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
                     /label="CMV promoter"
     promoter        678..696
                     /note="promoter for bacteriophage T7 RNA polymerase"
                     /label="T7 promoter"
     CDS             756..1577
                     /label="SLC25A26(NM_173471)"
                     /note="SLC25A26(NM_173471)"
                     /gene="SLC25A26"
     CDS             1584..1649
                     /codon_start=1
                     /product="three tandem FLAG® epitope tags, followed by an
                     enterokinase cleavage site"
                     /transl_table=1
                     /translation="DYKDHDGDYKDHDIDYKDDDDK"
                     /label="3xFLAG"
     misc_feature    1673..2261
                     /note="woodchuck hepatitis virus&nbsp;posttranscriptional
                     regulatory element"
                     /label="WPRE"
     polyA_signal    2327..2375
                     /note="herpes simplex virus thymidine kinase
                     polyadenylation signal&nbsp;(Cole and Stacy, 1985)"
                     /label="HSV TK poly(A) signal"
     rep_origin      2577..3005
                     /direction=RIGHT
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
                     /label="f1 ori"
     promoter        3019..3348
                     /note="SV40 enhancer and early promoter"
                     /label="SV40 promoter"
     CDS             3415..4209
                     /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    4385..4506
                     /note="SV40 polyadenylation signal"
                     /label="SV40 poly(A) signal"
     primer_bind     complement(4555..4571)
                     /note="common sequencing primer, one of multiple similar
                     variants"
                     /label="M13 rev"
     protein_bind    4579..4595
                     /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(4603..4633)
                     /note="promoter for the E. coli lac operon"
                     /label="lac promoter"
     protein_bind    4648..4669
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
                     /label="CAP binding site"
     rep_origin      complement(4957..5545)
                     /direction=LEFT
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of
                     replication"
                     /label="ori"
     CDS             complement(5716..6576)
                     /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(6577..6681)
                     /gene="bla"
                     /label="AmpR promoter"
ORIGIN
        1 gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgttg acattgatta
       61 ttgactagtt attaatagta atcaattacg gggtcattag ttcatagccc atatatggag
      121 ttccgcgtta cataacttac ggtaaatggc ccgcctggct gaccgcccaa cgacccccgc
      181 ccattgacgt caataatgac gtatgttccc atagtaacgc caatagggac tttccattga
      241 cgtcaatggg tggagtattt acggtaaact gcccacttgg cagtacatca agtgtatcat
      301 atgccaagta cgccccctat tgacgtcaat gacggtaaat ggcccgcctg gcattatgcc
      361 cagtacatga ccttatggga ctttcctact tggcagtaca tctacgtatt agtcatcgct
      421 attaccatgg tgatgcggtt ttggcagtac atcaatgggc gtggatagcg gtttgactca
      481 cggggatttc caagtctcca ccccattgac gtcaatggga gtttgttttg gcaccaaaat
      541 caacgggact ttccaaaatg tcgtaacaac tccgccccat tgacgcaaat gggcggtagg
      601 cgtgtacggt gggaggtcta tataagcaga gctctctggc taactagaga acccactgct
      661 tactggctta tcgaaattaa tacgactcac tatagggaga cccaagctgg ctagcgttta
      721 aacttaagct tggtaccgag ctcgaattcg ccaccatgga ccggccgggg ttcgtggcag
      781 cgctggtggc tggtggggta gcaggtgttt ctgttgactt gatattattt cctctggata
      841 ccattaaaac caggctgcag agtccccaag gatttagtaa ggctggtggt tttcatggaa
      901 tatatgctgg cgttccttct gctgctattg gatcctttcc taatgctgct gcatttttta
      961 tcacctatga atatgtgaag tggtttttgc atgctgattc atcttcatat ttgacaccta
     1021 tgaaacatat gttggctgcc tctgctggag aagtggttgc ctgcctgatt cgagttccat
     1081 ctgaagtggt taagcagagg gcacaggtat ctgcttctac aagaacattt cagattttct
     1141 ctaacatctt atatgaagag ggtatccaag ggttgtatcg aggctataaa agcacagttt
     1201 taagagagat tcctttttct ttggtccagt ttcccttatg ggagtcctta aaagccctct
     1261 ggtcctggag gcaggatcat gtggtggatt cttggcagtc agcagtctgt ggagcttttg
     1321 caggtggatt tgccgctgca gtcaccaccc ctctagacgt ggcaaagaca agaattacgc
     1381 tggcaaaggc tggctccagc actgctgatg ggaatgtgct ctctgtcctg catggggtct
     1441 ggcggtcaca ggggctggca ggattatttg caggtgtctt ccctcgaatg gcagccatca
     1501 gtctgggagg tttcatcttt ctgggggctt atgaccgaac gcacagcttg ctgttggaag
     1561 ttggcagaaa gagtcctgga tccgactaca aagaccatga cggtgattat aaagatcatg
     1621 acatcgacta caaggatgac gatgacaagt agtgagggcc cgatatctcg acaatcaacc
     1681 tctggattac aaaatttgtg aaagattgac tggtattctt aactatgttg ctccttttac
     1741 gctatgtgga tacgctgctt taatgccttt gtatcatgct attgcttccc gtatggcttt
     1801 cattttctcc tccttgtata aatcctggtt gctgtctctt tatgaggagt tgtggcccgt
     1861 tgtcaggcaa cgtggcgtgg tgtgcactgt gtttgctgac gcaaccccca ctggttgggg
     1921 cattgccacc acctgtcagc tcctttccgg gactttcgct ttccccctcc ctattgccac
     1981 ggcggaactc atcgccgcct gccttgcccg ctgctggaca ggggctcggc tgttgggcac
     2041 tgacaattcc gtggtgttgt cggggaagct gacgtccttt ccatggctgc tcgcctgtgt
     2101 tgccacctgg attctgcgcg ggacgtcctt ctgctacgtc ccttcggccc tcaatccagc
     2161 ggaccttcct tcccgcggcc tgctgccggc tctgcggcct cttccgcgtc ttcgccttcg
     2221 ccctcagacg agtcggatct ccctttgggc cgcctccccg cctggaaacg ggggaggcta
     2281 actgaaacac ggaaggagac aataccggaa ggaacccgcg ctatgacggc aataaaaaga
     2341 cagaataaaa cgcacgggtg ttgggtcgtt tgttcataaa cgcggggttc ggtcccaggg
     2401 ctggcactct gtcgataccc caccgagacc ccattggggc caatacgccc gcgtttcttc
     2461 cttttcccca ccccaccccc caagttcggg tgaaggccca gggctcgcag ccaacgtcgg
     2521 ggcggcaggc cctgccatag cagatctgcg cagctggggc tctagggggt atccccacgc
     2581 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
     2641 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
     2701 cgccggcttt ccccgtcaag ctctaaatcg gggcatccct ttagggttcc gatttagtgc
     2761 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
     2821 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
     2881 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
     2941 gattttgggg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
     3001 gaattaattc tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca
     3061 ggcagaagta tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca
     3121 ggctccccag caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc
     3181 ccgcccctaa ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc
     3241 catggctgac taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta
     3301 ttccagaagt agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctcccggga
     3361 gcttgtatat ccattttcgg atctgatcaa gagacaggat gaggatcgtt tcgcatgatt
     3421 gaacaagatg gattgcacgc aggttctccg gccgcttggg tggagaggct attcggctat
     3481 gactgggcac aacagacaat cggctgctct gatgccgccg tgttccggct gtcagcgcag
     3541 gggcgcccgg ttctttttgt caagaccgac ctgtccggtg ccctgaatga actgcaggac
     3601 gaggcagcgc ggctatcgtg gctggccacg acgggcgttc cttgcgcagc tgtgctcgac
     3661 gttgtcactg aagcgggaag ggactggctg ctattgggcg aagtgccggg gcaggatctc
     3721 ctgtcatctc accttgctcc tgccgagaaa gtatccatca tggctgatgc aatgcggcgg
     3781 ctgcatacgc ttgatccggc tacctgccca ttcgaccacc aagcgaaaca tcgcatcgag
     3841 cgagcacgta ctcggatgga agccggtctt gtcgatcagg atgatctgga cgaagagcat
     3901 caggggctcg cgccagccga actgttcgcc aggctcaagg cgcgcatgcc cgacggcgag
     3961 gatctcgtcg tgacccatgg cgatgcctgc ttgccgaata tcatggtgga aaatggccgc
     4021 ttttctggat tcatcgactg tggccggctg ggtgtggcgg accgctatca ggacatagcg
     4081 ttggctaccc gtgatattgc tgaagagctt ggcggcgaat gggctgaccg cttcctcgtg
     4141 ctttacggta tcgccgctcc cgattcgcag cgcatcgcct tctatcgcct tcttgacgag
     4201 ttcttctgag cgggactctg gggttcgcga aatgaccgac caagcgacgc ccaacctgcc
     4261 atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg gaatcgtttt
     4321 ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt tcttcgccca
     4381 ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca tcacaaattt
     4441 cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac tcatcaatgt
     4501 atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat catggtcata
     4561 gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac gagccggaag
     4621 cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa ttgcgttgcg
     4681 ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat gaatcggcca
     4741 acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc tcactgactc
     4801 gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg cggtaatacg
     4861 gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag gccagcaaaa
     4921 ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc gcccccctga
     4981 cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag gactataaag
     5041 ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga ccctgccgct
     5101 taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc aatgctcacg
     5161 ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg tgcacgaacc
     5221 ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt ccaacccggt
     5281 aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca gagcgaggta
     5341 tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca ctagaaggac
     5401 agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag ttggtagctc
     5461 ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca agcagcagat
     5521 tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg ggtctgacgc
     5581 tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa aaaggatctt
     5641 cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta tatatgagta
     5701 aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag cgatctgtct
     5761 atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga tacgggaggg
     5821 cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac cggctccaga
     5881 tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc ctgcaacttt
     5941 atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta gttcgccagt
     6001 taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac gctcgtcgtt
     6061 tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat gatcccccat
     6121 gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa gtaagttggc
     6181 cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg tcatgccatc
     6241 cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag aatagtgtat
     6301 gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc cacatagcag
     6361 aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct caaggatctt
     6421 accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat cttcagcatc
     6481 ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg ccgcaaaaaa
     6541 gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc aatattattg
     6601 aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta tttagaaaaa
     6661 taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg tcgacggatc
     6721 gggagatctc ccgatcccct atggtcgact ctcagtacaa tctgctctga tgccgcatag
     6781 ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg cgcgagcaaa
     6841 atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct gcttagg
//
