LOCUS       sequence-386564-        7055 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 7055)
  AUTHORS   Chou MY, Rooke N, Turck CW, Black DL
  TITLE     hnRNP H is a component of a splicing enhancer complex that activates
            a c-src alternative exon in neuronal cells.
  JOURNAL   Mol Cell Biol. 1999 Jan . 19(1):69-77.
  PUBMED    9858532
REFERENCE   2  (bases 1 to 7055)
  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: "Mol Cell 
            Biol. 1999 Jan . 19(1):69-77."
COMMENT     Sequence Label: pET15b-hnRNP-H
FEATURES             Location/Qualifiers
     source          1..7055
                     /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"
     protein_bind    20..44
                     /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)."
     RBS             59..81
                     /note="efficient ribosome binding site from bacteriophage 
                     T7 gene 10 (Olins and Rangwala, 1989)"
     CDS             100..117
                     /codon_start=1
                     /product="6xHis affinity tag"
                     /label=6xHis
                     /translation="HHHHHH"
     CDS             127..144
                     /codon_start=1
                     /product="thrombin recognition and cleavage site"
                     /label=thrombin site
                     /translation="LVPRGS"
     primer_bind     complement(1557..1575)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1571..1618
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(1793..1798,1799..1815,1816..1821))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(1848..1866)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        1934..2038
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(2039..2107,2108..2899)
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRVDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPAAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     complement(2257..2276)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      3070..3658
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3559..3578
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3812..3829
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    3844..3983
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(3909..3928)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     4069..4091
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(4085..4276)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     6631..6650
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(6670..6747)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     6953..6972
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggcagcagcc atcatcatca tcatcacagc
      121 agcggcctgg tgccgcgcgg cagccatatg ctcgagatga tgttgggcac ggagggtgga
      181 gagggattcg tggtgaaggt ccggggcttg ccctggtctt gctcggccga tgaagtgcag
      241 aggttttttt ctgactgcaa aattcaaaat ggggctcaag gtattcgttt catctacacc
      301 agagaaggca gaccaagtgg cgaggctttt gttgaacttg aatcagaaga tgaagtcaaa
      361 ttggccctga aaaaagacag agaaactatg ggacacagat atgttgaagt attcaagtca
      421 aacaacgttg aaatggattg ggtgttgaag catactggtc caaatagtcc tgacacggcc
      481 aatgatggct ttgtacggct tagaggactt ccctttggat gtagcaagga agaaattgtt
      541 cagttcttct cagggttgga aatcgtgcca aatgggataa cattgccggt ggacttccag
      601 gggaggagta cgggggaggc cttcgtgcag tttgcttcac aggaaatagc tgaaaaggct
      661 ctaaagaaac acaaggaaag aatagggcac aggtatattg aaatctttaa gagcagtaga
      721 gctgaagtta gaactcatta tgatccacca cgaaagctta tggccatgca gcggccaggt
      781 ccttatgaca gacctggggc tggtagaggg tataacagca ttggcagagg agctggcttt
      841 gagaggatga ggcgtggtgc ttatggtgga ggctatggag gctatgatga ttacaatggc
      901 tataatgatg gctatggatt tgggtcagat agatttggaa gagacctcaa ttactgtttt
      961 tcaggaatgt ctgatcacag atacggggat ggtggctcta ctttccagag cacaacagga
     1021 cactgtgtac acatgcgggg attaccttac agagctactg agaatgacat ttataatttt
     1081 ttttcaccgc tcaaccctgt gagagtacac attgaaattg gtcctgatgg cagagtaact
     1141 ggtgaagcag atgtcgagtt cgcaactcat gaagatgctg tggcagctat gtcaaaagac
     1201 aaagcaaata tgcaacacag atatgtagaa ctcttcttga attctacagc aggagcaagc
     1261 ggtggtgctt acgaacacag atatgtagaa ctcttcttga attctacagc aggagcaagc
     1321 ggtggtgctt atggtagcca aatgatggga ggcatgggct tgtcaaacca gtccagctac
     1381 gggggcccag ccagccagca gctgagtggg ggttacggag gcggctacgg tggccagagc
     1441 agcatgagtg gatacgacca agttttacag gaaaactcca gtgattttca atcaaacatt
     1501 gcatagggat ccggctgcta acaaagcccg aaaggaagct gagttggctg ctgccaccgc
     1561 tgagcaataa ctagcataac cccttggggc ctctaaacgg gtcttgaggg gttttttgct
     1621 gaaaggagga actatatccg gatatcccgc aagaggcccg gcagtaccgg cataaccaag
     1681 cctatgccta cagcatccag ggtgacggtg ccgaggatga cgatgagcgc attgttagat
     1741 ttcatacacg gtgcctgact gcgttagcaa tttaactgtg ataaactacc gcattaaagc
     1801 ttatcgatga taagctgtca aacatgagaa ttcttgaaga cgaaagggcc tcgtgatacg
     1861 cctattttta taggttaatg tcatgataat aatggtttct tagacgtcag gtggcacttt
     1921 tcggggaaat gtgcgcggaa cccctatttg tttatttttc taaatacatt caaatatgta
     1981 tccgctcatg agacaataac cctgataaat gcttcaataa tattgaaaaa ggaagagtat
     2041 gagtattcaa catttccgtg tcgcccttat tccctttttt gcggcatttt gccttcctgt
     2101 ttttgctcac ccagaaacgc tggtgaaagt aaaagatgct gaagatcagt tgggtgcacg
     2161 agtgggttac atcgaactgg atctcaacag cggtaagatc cttgagagtt ttcgccccga
     2221 agaacgtttt ccaatgatga gcacttttaa agttctgcta tgtggcgcgg tattatcccg
     2281 tgttgacgcc gggcaagagc aactcggtcg ccgcatacac tattctcaga atgacttggt
     2341 tgagtactca ccagtcacag aaaagcatct tacggatggc atgacagtaa gagaattatg
     2401 cagtgctgcc ataaccatga gtgataacac tgcggccaac ttacttctga caacgatcgg
     2461 aggaccgaag gagctaaccg cttttttgca caacatgggg gatcatgtaa ctcgccttga
     2521 tcgttgggaa ccggagctga atgaagccat accaaacgac gagcgtgaca ccacgatgcc
     2581 tgcagcaatg gcaacaacgt tgcgcaaact attaactggc gaactactta ctctagcttc
     2641 ccggcaacaa ttaatagact ggatggaggc ggataaagtt gcaggaccac ttctgcgctc
     2701 ggcccttccg gctggctggt ttattgctga taaatctgga gccggtgagc gtgggtctcg
     2761 cggtatcatt gcagcactgg ggccagatgg taagccctcc cgtatcgtag ttatctacac
     2821 gacggggagt caggcaacta tggatgaacg aaatagacag atcgctgaga taggtgcctc
     2881 actgattaag cattggtaac tgtcagacca agtttactca tatatacttt agattgattt
     2941 aaaacttcat ttttaattta aaaggatcta ggtgaagatc ctttttgata atctcatgac
     3001 caaaatccct taacgtgagt tttcgttcca ctgagcgtca gaccccgtag aaaagatcaa
     3061 aggatcttct tgagatcctt tttttctgcg cgtaatctgc tgcttgcaaa caaaaaaacc
     3121 accgctacca gcggtggttt gtttgccgga tcaagagcta ccaactcttt ttccgaaggt
     3181 aactggcttc agcagagcgc agataccaaa tactgtcctt ctagtgtagc cgtagttagg
     3241 ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa tcctgttacc
     3301 agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa gacgatagtt
     3361 accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc ccagcttgga
     3421 gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa gcgccacgct
     3481 tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa caggagagcg
     3541 cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg ggtttcgcca
     3601 cctctgactt gagcgtcgat ttttgtgatg ctcgtcaggg gggcggagcc tatggaaaaa
     3661 cgccagcaac gcggcctttt tacggttcct ggccttttgc tggccttttg ctcacatgtt
     3721 ctttcctgcg ttatcccctg attctgtgga taaccgtatt accgcctttg agtgagctga
     3781 taccgctcgc cgcagccgaa cgaccgagcg cagcgagtca gtgagcgagg aagcggaaga
     3841 gcgcctgatg cggtattttc tccttacgca tctgtgcggt atttcacacc gcaatggtgc
     3901 actctcagta caatctgctc tgatgccgca tagttaagcc agtatacact ccgctatcgc
     3961 tacgtgactg ggtcatggct gcgccccgac acccgccaac acccgctgac gcgccctgac
     4021 gggcttgtct gctcccggca tccgcttaca gacaagctgt gaccgtctcc gggagctgca
     4081 tgtgtcagag gttttcaccg tcatcaccga aacgcgcgag gcagctgcgg taaagctcat
     4141 cagcgtggtc gtgaagcgat tcacagatgt ctgcctgttc atccgcgtcc agctcgttga
     4201 gtttctccag aagcgttaat gtctggcttc tgataaagcg ggccatgtta agggcggttt
     4261 tttcctgttt ggtcactgat gcctccgtgt aagggggatt tctgttcatg ggggtaatga
     4321 taccgatgaa acgagagagg atgctcacga tacgggttac tgatgatgaa catgcccggt
     4381 tactggaacg ttgtgagggt aaacaactgg cggtatggat gcggcgggac cagagaaaaa
     4441 tcactcaggg tcaatgccag cgcttcgtta atacagatgt aggtgttcca cagggtagcc
     4501 agcagcatcc tgcgatgcag atccggaaca taatggtgca gggcgctgac ttccgcgttt
     4561 ccagacttta cgaaacacgg aaaccgaaga ccattcatgt tgttgctcag gtcgcagacg
     4621 ttttgcagca gcagtcgctt cacgttcgct cgcgtatcgg tgattcattc tgctaaccag
     4681 taaggcaacc ccgccagcct agccgggtcc tcaacgacag gagcacgatc atgcgcaccc
     4741 gtggccagga cccaacgctg cccgagatgc gccgcgtgcg gctgctggag atggcggacg
     4801 cgatggatat gttctgccaa gggttggttt gcgcattcac agttctccgc aagaattgat
     4861 tggctccaat tcttggagtg gtgaatccgt tagcgaggtg ccgccggctt ccattcaggt
     4921 cgaggtggcc cggctccatg caccgcgacg caacgcgggg aggcagacaa ggtatagggc
     4981 ggcgcctaca atccatgcca acccgttcca tgtgctcgcc gaggcggcat aaatcgccgt
     5041 gacgatcagc ggtccaatga tcgaagttag gctggtaaga gccgcgagcg atccttgaag
     5101 ctgtccctga tggtcgtcat ctacctgcct ggacagcatg gcctgcaacg cgggcatccc
     5161 gatgccgccg gaagcgagaa gaatcataat ggggaaggcc atccagcctc gcgtcgcgaa
     5221 cgccagcaag acgtagccca gcgcgtcggc cgccatgccg gcgataatgg cctgcttctc
     5281 gccgaaacgt ttggtggcgg gaccagtgac gaaggcttga gcgagggcgt gcaagattcc
     5341 gaataccgca agcgacaggc cgatcatcgt cgcgctccag cgaaagcggt cctcgccgaa
     5401 aatgacccag agcgctgccg gcacctgtcc tacgagttgc atgataaaga agacagtcat
     5461 aagtgcggcg acgatagtca tgccccgcgc ccaccggaag gagctgactg ggttgaaggc
     5521 tctcaagggc atcggtcgag atcccggtgc ctaatgagtg agctaactta cattaattgc
     5581 gttgcgctca ctgcccgctt tccagtcggg aaactgtcgt gccagctgca ttaatgaatc
     5641 ggccaacgcg cggggagagg cggtttgcgt attgggcgcc agggtggttt ttcttttcac
     5701 cagtgagacg ggcaacagct gattgccctt caccgcctgg ccctgagaga gttgcagcaa
     5761 gcggtccacg ctggtttgcc ccagcaggcg aaaatcctgt ttgatggtgg ttaacggcgg
     5821 gatataacat gagctgtctt cggtatcgtc gtatcccact accgagatat ccgcaccaac
     5881 gcgcagcccg gactcggtaa tggcgcgcat tgcgcccagc gccatctgat cgttggcaac
     5941 cagcatcgca gtgggaacga tgccctcatt cagcatttgc atggtttgtt gaaaaccgga
     6001 catggcactc cagtcgcctt cccgttccgc tatcggctga atttgattgc gagtgagata
     6061 tttatgccag ccagccagac gcagacgcgc cgagacagaa cttaatgggc ccgctaacag
     6121 cgcgatttgc tggtgaccca atgcgaccag atgctccacg cccagtcgcg taccgtcttc
     6181 atgggagaaa ataatactgt tgatgggtgt ctggtcagag acatcaagaa ataacgccgg
     6241 aacattagtg caggcagctt ccacagcaat ggcatcctgg tcatccagcg gatagttaat
     6301 gatcagccca ctgacgcgtt gcgcgagaag attgtgcacc gccgctttac aggcttcgac
     6361 gccgcttcgt tctaccatcg acaccaccac gctggcaccc agttgatcgg cgcgagattt
     6421 aatcgccgcg acaatttgcg acggcgcgtg cagggccaga ctggaggtgg caacgccaat
     6481 cagcaacgac tgtttgcccg ccagttgttg tgccacgcgg ttgggaatgt aattcagctc
     6541 cgccatcgcc gcttccactt tttcccgcgt tttcgcagaa acgtggctgg cctggttcac
     6601 cacgcgggaa acggtctgat aagagacacc ggcatactct gcgacatcgt ataacgttac
     6661 tggtttcaca ttcaccaccc tgaattgact ctcttccggg cgctatcatg ccataccgcg
     6721 aaaggttttg cgccattcga tggtgtccgg gatctcgacg ctctccctta tgcgactcct
     6781 gcattaggaa gcagcccagt agtaggttga ggccgttgag caccgccgcc gcaaggaatg
     6841 gtgcatgcaa ggagatggcg cccaacagtc ccccggccac ggggcctgcc accataccca
     6901 cgccgaaaca agcgctcatg agcccgaagt ggcgagcccg atcttcccca tcggtgatgt
     6961 cggcgatata ggcgccagca accgcacctg tggcgccggt gatgccggcc acgatgcgtc
     7021 cggcgtagag gatcgagatc tcgatcccgc gaaat
//
