LOCUS       sequence-266909-        8204 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 8204)
  AUTHORS   Morizono K, Xie Y, Ringpis GE, Johnson M, Nassanian H, Lee B, Wu L, 
            Chen IS
  TITLE     Lentiviral vector retargeting to P-glycoprotein on metastatic 
            melanoma through intravenous injection.
  JOURNAL   Nat Med. 2005 Mar;11(3):346-52. Epub 2005 Feb 13.
  PUBMED    15711560
REFERENCE   2  (bases 1 to 8204)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Dec 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "Nat Med."; 
            date: "2005-03"; volume: "11(3)"; pages: "346-52. Epub 2005 Feb 13"
COMMENT     Sequence Label: m168
FEATURES             Location/Qualifiers
     source          1..8204
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(35..52)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(206..794)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(286..305)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(965..1756,1757..1825))
                     /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     1588..1607
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(1826..1930)
                     /gene="bla"
                     /label=AmpR promoter
     rep_origin      complement(1956..2411)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2093..2114)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     2305..2324
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2538..2560
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     primer_bind     2552..2569
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     2553..2569
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2579..2598
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        2579..2597
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     enhancer        2718..3097
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        3098..3301
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     3252..3272
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     3298..3322
                     /label=LNCX
                     /note="Human CMV promoter, forward primer"
     intron          3413..3985
                     /label=beta-globin intron
                     /note="intron from rabbit beta-globin gene"
     CDS             4502..4672
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="DNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLLA
                     EAKKLNDAQAPK"
     CDS             4673..4846
                     /codon_start=1
                     /product="IgG-binding unit of Staphylococcus aureus protein
                     A"
                     /label=ProtA
                     /translation="VDNKFNKEQQNAFYEILHLPNLNEEQRNAFIQSLKDDPSQSANLL
                     AEAKKLNDAQAPK"
     primer_bind     complement(7530..7549)
                     /label=Bglob-pA-R
                     /note="Rabbit beta-globin polyA region, reverse primer"
     polyA_signal    7595..7650
                     /label=beta-globin poly(A) signal
                     /note="rabbit beta-globin polyadenylation signal (Gil and 
                     Proudfoot, 1987)"
     primer_bind     complement(7649..7668)
                     /label=rbglobpA-R
                     /note="Rabbit beta-globin polyA, reverse primer. Also 
                     called rb-glob-pA-term-R"
     primer_bind     complement(8008..8024)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(8008..8024)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(8021..8043)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    8032..8048
                     /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)."
     promoter        complement(join(8056..8062,8063..8080,8081..8086))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    8101..8122
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
ORIGIN
        1 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
       61 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
      121 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
      181 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
      241 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
      301 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
      361 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
      421 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
      481 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
      541 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
      601 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
      661 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
      721 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
      781 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
      841 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
      901 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
      961 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     1021 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     1081 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     1141 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     1201 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     1261 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     1321 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     1381 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     1441 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     1501 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     1561 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     1621 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     1681 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     1741 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     1801 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     1861 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     1921 gggttccgcg cacatttccc cgaaaagtgc cacctaaatt gtaagcgtta atattttgtt
     1981 aaaattcgcg ttaaattttt gttaaatcag ctcatttttt aaccaatagg ccgaaatcgg
     2041 caaaatccct tataaatcaa aagaatagac cgagataggg ttgagtgttg ttccagtttg
     2101 gaacaagagt ccactattaa agaacgtgga ctccaacgtc aaagggcgaa aaaccgtcta
     2161 tcagggcgat ggcccactac gtgaaccatc accctaatca agttttttgg ggtcgaggtg
     2221 ccgtaaagca ctaaatcgga accctaaagg gagcccccga tttagagctt gacggggaaa
     2281 gccggcgaac gtggcgagaa aggaagggaa gaaagcgaaa ggagcgggcg ctagggcgct
     2341 ggcaagtgta gcggtcacgc tgcgcgtaac caccacaccc gccgcgctta atgcgccgct
     2401 acagggcgcg tcccattcgc cattcaggct gcgcaactgt tgggaagggc gatcggtgcg
     2461 ggcctcttcg ctattacgcc agctggcgaa agggggatgt gctgcaaggc gattaagttg
     2521 ggtaacgcca gggttttccc agtcacgacg ttgtaaaacg acggccagtg agcgcgcgta
     2581 atacgactca ctatagggcg aattggagct ccaccgcggt ggcggccgct ctagagagct
     2641 tggcccattg catacgttgt atccatatca taatatgtac atttatattg gctcatgtcc
     2701 aacattaccg ccatgttgac attgattatt gactagttat taatagtaat caattacggg
     2761 gtcattagtt catagcccat atatggagtt ccgcgttaca taacttacgg taaatggccc
     2821 gcctggctga ccgcccaacg acccccgccc attgacgtca ataatgacgt atgttcccat
     2881 agtaacgcca atagggactt tccattgacg tcaatgggtg gagtatttac ggtaaactgc
     2941 ccacttggca gtacatcaag tgtatcatat gccaagtacg ccccctattg acgtcaatga
     3001 cggtaaatgg cccgcctggc attatgccca gtacatgacc ttatgggact ttcctacttg
     3061 gcagtacatc tacgtattag tcatcgctat taccatggtg atgcggtttt ggcagtacat
     3121 caatgggcgt ggatagcggt ttgactcacg gggatttcca agtctccacc ccattgacgt
     3181 caatgggagt ttgttttggc accaaaatca acgggacttt ccaaaatgtc gtaacaactc
     3241 cgccccattg acgcaaatgg gcggtaggcg tgtacggtgg gaggtctata taagcagagc
     3301 tcgtttagtg aaccgtcaga tcgcctggag acgccatcca cgctgttttg acctccatag
     3361 aagacaccgg gaccgatcca gcctccggtc gaccgatcct gagaacttca gggtgagttt
     3421 ggggaccctt gattgttctt tctttttcgc tattgtaaaa ttcatgttat atggaggggg
     3481 caaagttttc agggtgttgt ttagaatggg aagatgtccc ttgtatcacc atggaccctc
     3541 atgataattt tgtttctttc actttctact ctgttgacaa ccattgtctc ctcttatttt
     3601 cttttcattt tctgtaactt tttcgttaaa ctttagcttg catttgtaac gaatttttaa
     3661 attcactttt gtttatttgt cagattgtaa gtactttctc taatcacttt tttttcaagg
     3721 caatcagggt atattatatt gtacttcagc acagttttag agaacaattg ttataattaa
     3781 atgataaggt agaatatttc tgcatataaa ttctggctgg cgtggaaata ttcttattgg
     3841 tagaaacaac tacaccctgg tcatcatcct gcctttctct ttatggttac aatgatatac
     3901 actgtttgag atgaggataa aatactctga gtccaaaccg ggcccctctg ctaaccatgt
     3961 tcatgccttc ttctctttcc tacagctcct gggcaacgtg ctggttgttg tgctgtctca
     4021 tcattttggc aaagaattcc tcgacggatc cctcgaccta gacatttcat ctgactaata
     4081 ctacaacacc accaccatgg cgtccgcagc accactggtc acggcaatgt gtttgctcgg
     4141 aaatgtgagc ttcccatgcg accgcccgcc cacatgctat acccgcgaac cttccagagc
     4201 cctcgacatc cttgaagaga acgtgaacca tgaggcctac gataccctgc tcaatgccat
     4261 attgcggtgc ggatcgtctg gcagcgtcat tgacgacttt accctgacca gcccctactt
     4321 gggcacatgc tcgtactgcc accatactga accgtgcttc agccctgtta agatcgagca
     4381 ggtctgggac gaagcggacg ataacaccat acgcatacag acttccgccc agtttggata
     4441 cgaccaaagc ggagcagcaa gcgcaaacaa gtaccgctac atggcggctg cggcggtaac
     4501 cgacaacaaa ttcaacaaag aacaacaaaa cgcgttctat gagatcttac atttacctaa
     4561 cttaaacgaa gaacaacgaa acgccttcat ccaaagttta aaagatgacc caagccaaag
     4621 cgctaacctt ttagcagaag ctaaaaagct aaatgatgct caggcgccga aagtagacaa
     4681 caaattcaac aaagaacaac aaaacgcgtt ctatgagatc ttacatttac ctaacttaaa
     4741 cgaagaacaa cgaaacgcct tcatccaaag tttaaaagat gacccaagcc aaagcgctaa
     4801 ccttttagca gaagctaaaa agctaaatga tgctcaggcg ccgaaagtag acgcgaattc
     4861 gagctcggta cccggggatc cggtaaccac cgttaaagaa ggcaccatgg atgacatcaa
     4921 gattagcacc tcaggaccgt gtagaaggct tagctacaaa ggatactttc tcctcgcaaa
     4981 atgccctcca ggggacagcg taacggttag catagtgagt agcaactcag caacgtcatg
     5041 tacactggcc cgcaagataa aaccaaaatt cgtgggacgg gaaaaatatg atctacctcc
     5101 cgttcacggt aaaaaaattc cttgcacagt gtacgaccgt ctggcagcaa caactgcagg
     5161 ctacatcact atgcacaggc cgggaccgca cgcttataca tcctacctgg aagaatcatc
     5221 agggaaagtt tacgcaaagc cgccatctgg gaagaacatt acgtatgagt gcaagtgcgg
     5281 cgactacaag accggaaccg tttcgacccg caccgaaatc actggttgca ccgccatcaa
     5341 gcagtgcgtc gcctataaga gcgaccaaac gaagtgggtc ttcaactcac cggacttgat
     5401 cagacatgac gaccacacgg tccaagggaa attgcatttg cctttcaagt tgatcccgag
     5461 tacctgcatg gtccctgttg cccacgcgcc gaatgtaata catggcttta aacacatcag
     5521 cctccaatta gatacagacc acttgacatt gctcaccacc aggagactag gggcaaaccc
     5581 ggaaccaacc actgaatgga tcgtcggaaa gacggtcaga aacttcaccg tcgaccgaga
     5641 tggcctggaa tacatatggg gaaatcatga gccagtgagg gtctatgccc aagagtcagc
     5701 accaggagac cctcacggat ggccacacga aatagtacag cattactacc atcgccatcc
     5761 tgtgtacacc atcttagccg tcgcatcagc taccgtggcg atgatgattg gcgtaactgt
     5821 tgcagtgtta tgtgcctgta aagcgcgccg tgagtgcctg acgccatacg ccctggcccc
     5881 aaacgccgta atcccaactt cgctggcact cttgtgctgc gttaggtcgg ccaatgctga
     5941 aacgttcacc gagaccatga gttacttgtg gtcgaacagt cagccgttct tctgggtcca
     6001 gttgtgcata cctttggccg ctttcatcgt tctaatgcgc tgctgctcct gctgcctgcc
     6061 ttttttagtg gttgccggcg cctacctggc gaaggtagac gcctacgaac atgcgaccac
     6121 tgttccaaat gtgccacaga taccgtataa ggcacttgtt gaaagggcag ggtatgcccc
     6181 gctcaatttg gagatcactg tcatgtcctc ggaggttttg ccttccacca accaagagta
     6241 cattacctgc aaattcacca ctgtggtccc ctccccaaaa atcaaatgct gcggctcctt
     6301 ggaatgtcag ccggccgctc atgcaggcta tacctgcaag gtcttcggag gggtctaccc
     6361 ctttatgtgg ggaggagcgc aatgtttttg cgacagtgag aacagccaga tgagtgaggc
     6421 gtacgtcgaa ttgtcagcag attgcgcgtc tgaccacgcg caggcgatta aggtgcacac
     6481 tgccgcgatg aaagtaggac tgcgtattgt gtacgggaac actaccagtt tcctagatgt
     6541 gtacgtgaac ggagtcacac caggaacgtc taaagacttg aaagtcatag ctggaccaat
     6601 ttcagcatcg tttacgccat tcgatcataa ggtcgttatc catcgcggcc tggtgtacaa
     6661 ctatgacttc ccggaatatg gagcgatgaa accaggagcg tttggagaca ttcaagctac
     6721 ctccttgact agcaaggatc tcatcgccag cacagacatt aggctactca agccttccgc
     6781 caagaacgtg catgtcccgt acacgcaggc ctcatcagga tttgagatgt ggaaaaacaa
     6841 ctcaggccgc ccactgcagg aaaccgcacc tttcgggtgt aagattgcag taaatccgct
     6901 ccgagcggtg gactgttcat acgggaacat tcccatttct attgacatcc cgaacgctgc
     6961 ctttatcagg acatcagatg caccactggt ctcaacagtc aaatgtgaag tcagtgagtg
     7021 cacttattca gcagacttcg gcgggatggc caccctgcag tatgtatccg accgcgaagg
     7081 tcaatgcccc gtacattcgc attcgagcac agcaactctc caagagtcga cagtacatgt
     7141 cctggagaaa ggagcggtga cagtacactt tagcaccgcg agtccacagg cgaactttat
     7201 cgtatcgctg tgtgggaaga agacaacatg caatgcagaa tgtaaaccac cagctgacca
     7261 tatcgtgagc accccgcaca aaaatgacca agaatttcaa gccgccatct caaaaacatc
     7321 atggagttgg ctgtttgccc ttttcggcgg cgcctcgtcg ctattaatta taggacttat
     7381 gatttttgct tgcagcatga tgctgactag cacacgaaga tgactcgagt cgagggatcc
     7441 gtcgaggaat tcactcctca ggtgcaggct gcctatcaga aggtggtggc tggtgtggcc
     7501 aatgccctgg ctcacaaata ccactgagat ctttttccct ctgccaaaaa ttatggggac
     7561 atcatgaagc cccttgagca tctgacttct ggctaataaa ggaaatttat tttcattgca
     7621 atagtgtgtt ggaatttttt gtgtctctca ctcggaagga catatgggag ggcaaatcat
     7681 ttaaaacatc agaatgagta tttggtttag agtttggcaa catatgccca tatgctggct
     7741 gccatgaaca aaggttggct ataaagaggt catcagtata tgaaacagcc ccctgctgtc
     7801 cattccttat tccatagaaa agccttgact tgaggttaga ttttttttat attttgtttt
     7861 gtgttatttt tttctttaac atccctaaaa ttttccttac atgttttact agccagattt
     7921 ttcctcctct cctgactact cccagtcata gctgtccctc ttctcttatg gagatccctc
     7981 gacggatcgg ccgcaattcg taatcatgtc atagctgttt cctgtgtgaa attgttatcc
     8041 gctcacaatt ccacacaaca tacgagccgg aagcataaag tgtaaagcct ggggtgccta
     8101 atgagtgagc taactcacat taattgcgtt gcgctcactg cccgctttcc agtcgggaaa
     8161 cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg ggga
//
