LOCUS       sequence-175674-        5777 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Bacterial expression of fusion protein with cell penetrating 
            peptide, antennapedia fragment, at C-terminus..
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5777)
  AUTHORS   Chien WM, Liu Y, Chin MT
  TITLE     Genomic DNA recombination with cell-penetrating peptide-tagged cre 
            protein in mouse skeletal and cardiac muscle.
  JOURNAL   Genesis. 2014 Jul;52(7):695-701. doi: 10.1002/dvg.22782. Epub 2014 
            May 5.
  PUBMED    24753043
REFERENCE   2  (bases 1 to 5777)
  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"; doi: "10.1002/dvg.22782";
            journalName: "Genesis"; date: "2014-07"; volume: "52"; issue: "7"; 
            pages: "695-701"
COMMENT     Sequence Label: pET16b-Antp
FEATURES             Location/Qualifiers
     source          1..5777
                     /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             94..123
                     /codon_start=1
                     /product="10xHis affinity tag"
                     /label=10xHis
                     /translation="HHHHHHHHHH"
     CDS             136..147
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
     primer_bind     complement(279..297)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      293..340
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     promoter        complement(join(515..520,521..537,538..543))
                     /label=tet promoter
                     /note="E. coli promoter for tetracycline efflux protein 
                     gene"
     primer_bind     complement(570..588)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     promoter        656..760
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(761..829,830..1621)
                     /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(979..998)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      1792..2380
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     2281..2300
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     2534..2551
                     /label=L4440
                     /note="L4440 vector, forward primer"
     misc_feature    2566..2705
                     /label=bom
                     /note="basis of mobility region from pBR322"
     primer_bind     complement(2631..2650)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     primer_bind     2791..2813
                     /label=pGEX 3'
                     /note="pGEX vectors, reverse primer"
     CDS             complement(2807..2998)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     primer_bind     5353..5372
                     /label=LacI-R
                     /note="LacI, reverse primer"
     promoter        complement(5392..5469)
                     /gene="lacI"
                     /label=lacI promoter
     primer_bind     5675..5694
                     /label=pBRrevBam
                     /note="pBR322 vectors, tet region, downstream of BamHI, 
                     reverse primer"
ORIGIN
        1 taatacgact cactataggg gaattgtgag cggataacaa ttcccctcta gaaataattt
       61 tgtttaactt taagaaggag atataccatg ggccatcatc atcatcatca tcatcatcat
      121 cacagcagcg gccatatcga aggtcgtcat atgctcgagg agtccggcgg aggcggatct
      181 cgccagataa agatttggtt ccagaatcgg cgcatgaagt ggaagaaggg atccggctgc
      241 taacaaagcc cgaaaggaag ctgagttggc tgctgccacc gctgagcaat aactagcata
      301 accccttggg gcctctaaac gggtcttgag gggttttttg ctgaaaggag gaactatatc
      361 cggatatccc gcaagaggcc cggcagtacc ggcataacca agcctatgcc tacagcatcc
      421 agggtgacgg tgccgaggat gacgatgagc gcattgttag atttcataca cggtgcctga
      481 ctgcgttagc aatttaactg tgataaacta ccgcattaaa gcttatcgat gataagctgt
      541 caaacatgag aattcttgaa gacgaaaggg cctcgtgata cgcctatttt tataggttaa
      601 tgtcatgata ataatggttt cttagacgtc aggtggcact tttcggggaa atgtgcgcgg
      661 aacccctatt tgtttatttt tctaaataca ttcaaatatg tatccgctca tgagacaata
      721 accctgataa atgcttcaat aatattgaaa aaggaagagt atgagtattc aacatttccg
      781 tgtcgccctt attccctttt ttgcggcatt ttgccttcct gtttttgctc acccagaaac
      841 gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca cgagtgggtt acatcgaact
      901 ggatctcaac agcggtaaga tccttgagag ttttcgcccc gaagaacgtt ttccaatgat
      961 gagcactttt aaagttctgc tatgtggcgc ggtattatcc cgtgttgacg ccgggcaaga
     1021 gcaactcggt cgccgcatac actattctca gaatgacttg gttgagtact caccagtcac
     1081 agaaaagcat cttacggatg gcatgacagt aagagaatta tgcagtgctg ccataaccat
     1141 gagtgataac actgcggcca acttacttct gacaacgatc ggaggaccga aggagctaac
     1201 cgcttttttg cacaacatgg gggatcatgt aactcgcctt gatcgttggg aaccggagct
     1261 gaatgaagcc ataccaaacg acgagcgtga caccacgatg cctgcagcaa tggcaacaac
     1321 gttgcgcaaa ctattaactg gcgaactact tactctagct tcccggcaac aattaataga
     1381 ctggatggag gcggataaag ttgcaggacc acttctgcgc tcggcccttc cggctggctg
     1441 gtttattgct gataaatctg gagccggtga gcgtgggtct cgcggtatca ttgcagcact
     1501 ggggccagat ggtaagccct cccgtatcgt agttatctac acgacgggga gtcaggcaac
     1561 tatggatgaa cgaaatagac agatcgctga gataggtgcc tcactgatta agcattggta
     1621 actgtcagac caagtttact catatatact ttagattgat ttaaaacttc atttttaatt
     1681 taaaaggatc taggtgaaga tcctttttga taatctcatg accaaaatcc cttaacgtga
     1741 gttttcgttc cactgagcgt cagaccccgt agaaaagatc aaaggatctt cttgagatcc
     1801 tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa ccaccgctac cagcggtggt
     1861 ttgtttgccg gatcaagagc taccaactct ttttccgaag gtaactggct tcagcagagc
     1921 gcagatacca aatactgtcc ttctagtgta gccgtagtta ggccaccact tcaagaactc
     1981 tgtagcaccg cctacatacc tcgctctgct aatcctgtta ccagtggctg ctgccagtgg
     2041 cgataagtcg tgtcttaccg ggttggactc aagacgatag ttaccggata aggcgcagcg
     2101 gtcgggctga acggggggtt cgtgcacaca gcccagcttg gagcgaacga cctacaccga
     2161 actgagatac ctacagcgtg agctatgaga aagcgccacg cttcccgaag ggagaaaggc
     2221 ggacaggtat ccggtaagcg gcagggtcgg aacaggagag cgcacgaggg agcttccagg
     2281 gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc cacctctgac ttgagcgtcg
     2341 atttttgtga tgctcgtcag gggggcggag cctatggaaa aacgccagca acgcggcctt
     2401 tttacggttc ctggcctttt gctggccttt tgctcacatg ttctttcctg cgttatcccc
     2461 tgattctgtg gataaccgta ttaccgcctt tgagtgagct gataccgctc gccgcagccg
     2521 aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa gagcgcctga tgcggtattt
     2581 tctccttacg catctgtgcg gtatttcaca ccgcaatggt gcactctcag tacaatctgc
     2641 tctgatgccg catagttaag ccagtataca ctccgctatc gctacgtgac tgggtcatgg
     2701 ctgcgccccg acacccgcca acacccgctg acgcgccctg acgggcttgt ctgctcccgg
     2761 catccgctta cagacaagct gtgaccgtct ccgggagctg catgtgtcag aggttttcac
     2821 cgtcatcacc gaaacgcgcg aggcagctgc ggtaaagctc atcagcgtgg tcgtgaagcg
     2881 attcacagat gtctgcctgt tcatccgcgt ccagctcgtt gagtttctcc agaagcgtta
     2941 atgtctggct tctgataaag cgggccatgt taagggcggt tttttcctgt ttggtcactg
     3001 atgcctccgt gtaaggggga tttctgttca tgggggtaat gataccgatg aaacgagaga
     3061 ggatgctcac gatacgggtt actgatgatg aacatgcccg gttactggaa cgttgtgagg
     3121 gtaaacaact ggcggtatgg atgcggcggg accagagaaa aatcactcag ggtcaatgcc
     3181 agcgcttcgt taatacagat gtaggtgttc cacagggtag ccagcagcat cctgcgatgc
     3241 agatccggaa cataatggtg cagggcgctg acttccgcgt ttccagactt tacgaaacac
     3301 ggaaaccgaa gaccattcat gttgttgctc aggtcgcaga cgttttgcag cagcagtcgc
     3361 ttcacgttcg ctcgcgtatc ggtgattcat tctgctaacc agtaaggcaa ccccgccagc
     3421 ctagccgggt cctcaacgac aggagcacga tcatgcgcac ccgtggccag gacccaacgc
     3481 tgcccgagat gcgccgcgtg cggctgctgg agatggcgga cgcgatggat atgttctgcc
     3541 aagggttggt ttgcgcattc acagttctcc gcaagaattg attggctcca attcttggag
     3601 tggtgaatcc gttagcgagg tgccgccggc ttccattcag gtcgaggtgg cccggctcca
     3661 tgcaccgcga cgcaacgcgg ggaggcagac aaggtatagg gcggcgccta caatccatgc
     3721 caacccgttc catgtgctcg ccgaggcggc ataaatcgcc gtgacgatca gcggtccaat
     3781 gatcgaagtt aggctggtaa gagccgcgag cgatccttga agctgtccct gatggtcgtc
     3841 atctacctgc ctggacagca tggcctgcaa cgcgggcatc ccgatgccgc cggaagcgag
     3901 aagaatcata atggggaagg ccatccagcc tcgcgtcgcg aacgccagca agacgtagcc
     3961 cagcgcgtcg gccgccatgc cggcgataat ggcctgcttc tcgccgaaac gtttggtggc
     4021 gggaccagtg acgaaggctt gagcgagggc gtgcaagatt ccgaataccg caagcgacag
     4081 gccgatcatc gtcgcgctcc agcgaaagcg gtcctcgccg aaaatgaccc agagcgctgc
     4141 cggcacctgt cctacgagtt gcatgataaa gaagacagtc ataagtgcgg cgacgatagt
     4201 catgccccgc gcccaccgga aggagctgac tgggttgaag gctctcaagg gcatcggtcg
     4261 agatcccggt gcctaatgag tgagctaact tacattaatt gcgttgcgct cactgcccgc
     4321 tttccagtcg ggaaactgtc gtgccagctg cattaatgaa tcggccaacg cgcggggaga
     4381 ggcggtttgc gtattgggcg ccagggtggt ttttcttttc accagtgaga cgggcaacag
     4441 ctgattgccc ttcaccgcct ggccctgaga gagttgcagc aagcggtcca cgctggtttg
     4501 ccccagcagg cgaaaatcct gtttgatggt ggttaacggc gggatataac atgagctgtc
     4561 ttcggtatcg tcgtatccca ctaccgagat atccgcacca acgcgcagcc cggactcggt
     4621 aatggcgcgc attgcgccca gcgccatctg atcgttggca accagcatcg cagtgggaac
     4681 gatgccctca ttcagcattt gcatggtttg ttgaaaaccg gacatggcac tccagtcgcc
     4741 ttcccgttcc gctatcggct gaatttgatt gcgagtgaga tatttatgcc agccagccag
     4801 acgcagacgc gccgagacag aacttaatgg gcccgctaac agcgcgattt gctggtgacc
     4861 caatgcgacc agatgctcca cgcccagtcg cgtaccgtct tcatgggaga aaataatact
     4921 gttgatgggt gtctggtcag agacatcaag aaataacgcc ggaacattag tgcaggcagc
     4981 ttccacagca atggcatcct ggtcatccag cggatagtta atgatcagcc cactgacgcg
     5041 ttgcgcgaga agattgtgca ccgccgcttt acaggcttcg acgccgcttc gttctaccat
     5101 cgacaccacc acgctggcac ccagttgatc ggcgcgagat ttaatcgccg cgacaatttg
     5161 cgacggcgcg tgcagggcca gactggaggt ggcaacgcca atcagcaacg actgtttgcc
     5221 cgccagttgt tgtgccacgc ggttgggaat gtaattcagc tccgccatcg ccgcttccac
     5281 tttttcccgc gttttcgcag aaacgtggct ggcctggttc accacgcggg aaacggtctg
     5341 ataagagaca ccggcatact ctgcgacatc gtataacgtt actggtttca cattcaccac
     5401 cctgaattga ctctcttccg ggcgctatca tgccataccg cgaaaggttt tgcgccattc
     5461 gatggtgtcc gggatctcga cgctctccct tatgcgactc ctgcattagg aagcagccca
     5521 gtagtaggtt gaggccgttg agcaccgccg ccgcaaggaa tggtgcatgc aaggagatgg
     5581 cgcccaacag tcccccggcc acggggcctg ccaccatacc cacgccgaaa caagcgctca
     5641 tgagcccgaa gtggcgagcc cgatcttccc catcggtgat gtcggcgata taggcgccag
     5701 caaccgcacc tgtggcgccg gtgatgccgg ccacgatgcg tccggcgtag aggatcgaga
     5761 tctcgatccc gcgaaat
//
