LOCUS       sequence-338155-        2997 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 2997)
  AUTHORS   Nagahara H, Vocero-Akbani AM, Snyder EL, Ho A, Latham DG, Lissy NA, 
            Becker-Hapak M, Ezhevsky SA, Dowdy SF
  TITLE     Transduction of full-length TAT fusion proteins into mammalian 
            cells: TAT-p27Kip1 induces cell migration.
  JOURNAL   Nat Med. 1998 Dec;4(12):1449-52.
  PUBMED    9846587
REFERENCE   2  (bases 1 to 2997)
  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: "1998-12"; volume: "4(12)"; pages: "1449-52"
COMMENT     Sequence Label: pTAT-HA
FEATURES             Location/Qualifiers
     source          1..2997
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     1..20
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        1..19
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     RBS             51..73
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             93..110
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             114..146
                     /codon_start=1
                     /product="leader peptide from bacteriophage T7 gene 10"
                     /label=T7 tag (gene 10 leader)
                     /note="promotes efficient translation in E. coli"
                     /translation="MASMTGGQQMG"
     CDS             join(150..158,159..173)
                     /codon_start=1
                     /product="Xpress(TM) epitope tag, including an enterokinase
                     recognition and cleavage site"
                     /label=Xpress(TM) tag
                     /translation="DLYDDDDK"
     CDS             252..278
                     /codon_start=1
                     /product="HA (human influenza hemagglutinin) epitope tag"
                     /label=HA
                     /translation="YPYDVPDYA"
     primer_bind     complement(379..397)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      393..440
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     rep_origin      537..992
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(624..643)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     834..855
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        1018..1122
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1123..1191,1192..1983)
                     /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(1341..1360)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2154..2742
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2643..2662
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2896..2913
                     /label=L4440
                     /note="L4440 vector, forward primer"
ORIGIN
        1 taatacgact cactataggg agaccacaac ggtttccctc tagaaataat tttgtttaac
       61 tttaagaagg agatatacat atgcggggtt ctcatcatca tcatcatcat ggtatggcta
      121 gcatgactgg tggacagcaa atgggtcggg atctgtacga cgatgacgat aaggatcgat
      181 ggggatccaa gcttggctac ggccgcaaga aacgccgcca gcgccgccgc ggtggatcca
      241 ccatgtccgg ctatccatat gacgtcccag actatgctgg ctccatggcc ggtaccggtc
      301 tcgaggtgca tgcggtgaat tcgaagcttg atccggctgc taacaaagcc cgaaaggaag
      361 ctgagttggc tgctgccacc gctgagcaat aactagcata accccttggg gcctctaaac
      421 gggtcttgag gggttttttg ctgaaaggag gaactatatc cggatctggc gtaatagcga
      481 agaggcccgc accgatcgcc cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc
      541 gccctgtagc ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac
      601 acttgccagc gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt
      661 cgccggcttt ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc
      721 tttacggcac ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc
      781 gccctgatag acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact
      841 cttgttccaa actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg
      901 gattttgccg atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc
      961 gaattttaac aaaatattaa cgcttacaat ttaggtggca cttttcgggg aaatgtgcgc
     1021 ggaaccccta tttgtttatt tttctaaata cattcaaata tgtatccgct catgagacaa
     1081 taaccctgat aaatgcttca ataatattga aaaaggaaga gtatgagtat tcaacatttc
     1141 cgtgtcgccc ttattccctt ttttgcggca ttttgccttc ctgtttttgc tcacccagaa
     1201 acgctggtga aagtaaaaga tgctgaagat cagttgggtg cacgagtggg ttacatcgaa
     1261 ctggatctca acagcggtaa gatccttgag agttttcgcc ccgaagaacg ttttccaatg
     1321 atgagcactt ttaaagttct gctatgtggc gcggtattat cccgtattga cgccgggcaa
     1381 gagcaactcg gtcgccgcat acactattct cagaatgact tggttgagta ctcaccagtc
     1441 acagaaaagc atcttacgga tggcatgaca gtaagagaat tatgcagtgc tgccataacc
     1501 atgagtgata acactgcggc caacttactt ctgacaacga tcggaggacc gaaggagcta
     1561 accgcttttt tgcacaacat gggggatcat gtaactcgcc ttgatcgttg ggaaccggag
     1621 ctgaatgaag ccataccaaa cgacgagcgt gacaccacga tgcctgtagc aatggcaaca
     1681 acgttgcgca aactattaac tggcgaacta cttactctag cttcccggca acaattaata
     1741 gactggatgg aggcggataa agttgcagga ccacttctgc gctcggccct tccggctggc
     1801 tggtttattg ctgataaatc tggagccggt gagcgtgggt ctcgcggtat cattgcagca
     1861 ctggggccag atggtaagcc ctcccgtatc gtagttatct acacgacggg gagtcaggca
     1921 actatggatg aacgaaatag acagatcgct gagataggtg cctcactgat taagcattgg
     1981 taactgtcag accaagttta ctcatatata ctttagattg atttaaaact tcatttttaa
     2041 tttaaaagga tctaggtgaa gatccttttt gataatctca tgaccaaaat cccttaacgt
     2101 gagttttcgt tccactgagc gtcagacccc gtagaaaaga tcaaaggatc ttcttgagat
     2161 cctttttttc tgcgcgtaat ctgctgcttg caaacaaaaa aaccaccgct accagcggtg
     2221 gtttgtttgc cggatcaaga gctaccaact ctttttccga aggtaactgg cttcagcaga
     2281 gcgcagatac caaatactgt tcttctagtg tagccgtagt taggccacca cttcaagaac
     2341 tctgtagcac cgcctacata cctcgctctg ctaatcctgt taccagtggc tgctgccagt
     2401 ggcgataagt cgtgtcttac cgggttggac tcaagacgat agttaccgga taaggcgcag
     2461 cggtcgggct gaacgggggg ttcgtgcaca cagcccagct tggagcgaac gacctacacc
     2521 gaactgagat acctacagcg tgagctatga gaaagcgcca cgcttcccga agggagaaag
     2581 gcggacaggt atccggtaag cggcagggtc ggaacaggag agcgcacgag ggagcttcca
     2641 gggggaaacg cctggtatct ttatagtcct gtcgggtttc gccacctctg acttgagcgt
     2701 cgatttttgt gatgctcgtc aggggggcgg agcctatgga aaaacgccag caacgcggcc
     2761 tttttacggt tcctggcctt ttgctggcct tttgctcaca tgttctttcc tgcgttatcc
     2821 cctgattctg tggataaccg tattaccgcc tttgagtgag ctgataccgc tcgccgcagc
     2881 cgaacgaccg agcgcagcga gtcagtgagc gaggaagcgg aagagcgccc aatacgcaaa
     2941 ccgcctctcc ccgcgcgttg gccgattcat taatgcagga tctcgatccc gcgaaat
//
