LOCUS       sequence-462334-        5026 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  lvl 1 backbone.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5026)
  AUTHORS   Duportet X, Wroblewska L, Guye P, Li Y, Eyquem J, Rieders J, 
            Rimchala T, Batt G, Weiss R
  TITLE     A platform for rapid prototyping of synthetic gene networks in 
            mammalian cells.
  JOURNAL   Nucleic Acids Res. 2014 Dec 1;42(21):13440-51. doi: 
            10.1093/nar/gku1082. Epub 2014 Nov 5.
  PUBMED    25378321
REFERENCE   2  (bases 1 to 5026)
  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: 
            "10.1093/nar/gku1082"; journalName: "Nucleic Acids Res"; date: 
            "2014-12-1- 1"; volume: "42"; issue: "21"; pages: "13440-51"
COMMENT     Sequence Label: lvl2-[piggyBac-backbone]
FEATURES             Location/Qualifiers
     source          1..5026
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     rep_origin      1..589
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     490..509
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     743..760
                     /label=L4440
                     /note="L4440 vector, forward primer"
     repeat_region   791..853
                     /label=piggyBac right (3') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     terminator      complement(1792..1835)
                     /label=bacterial terminator
                     /note="putative bacterial transcription terminator"
     misc_feature    1838..1859
                     /label=BioBrick prefix
                     /note="BioBrick prefix for parts that do not start with 
                     ""ATG"""
     protein_bind    1860..1878
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     protein_bind    1885..1903
                     /gene="tetO"
                     /label=tet operator
                     /bound_moiety="tetracycline repressor TetR"
                     /note="bacterial operator O2 for the tetR and tetA genes"
     RBS             1922..1933
                     /note="strong bacterial ribosome binding site (Elowitz and 
                     Leibler, 2000)"
     CDS             1940..2617
                     /codon_start=1
                     /product="monomeric derivative of DsRed (Campbell et al., 
                     2002)"
                     /label=mRFP1
                     /translation="MASSEDVIKEFMRFKVRMEGSVNGHEFEIEGEGEGRPYEGTQTAK
                     LKVTKGGPLPFAWDILSPQFQYGSKAYVKHPADIPDYLKLSFPEGFKWERVMNFEDGGV
                     VTVTQDSSLQDGEFIYKVKLRGTNFPSDGPVMQKKTMGWEASTERMYPEDGALKGEIKM
                     RLKLKDGGHYDAEVKTTYMAKKPVQLPGAYKTDIKLDITSHNEDYTIVEQYERAEGRHS
                     TGA"
     terminator      2662..2733
                     /label=rrnB T1 terminator
                     /note="transcription terminator T1 from the E. coli rrnB 
                     gene"
     terminator      2749..2776
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     repeat_region   3801..3835
                     /label=piggyBac left (5') inverted repeat
                     /note="piggyBac transposon-specific inverted terminal 
                     repeat sequence (ITR)"
     primer_bind     complement(4052..4071)
                     /label=pENTR-R
                     /note="pENTR vectors, reverse primer"
     CDS             4063..4872
                     /codon_start=1
                     /gene="aph(3')-Ia"
                     /product="aminoglycoside phosphotransferase"
                     /label=KanR
                     /note="confers resistance to kanamycin in bacteria or G418 
                     (Geneticin(R)) in eukaryotes"
                     /translation="MSHIQRETSRPRLNSNMDADLYGYKWARDNVGQSGATIYRLYGKP
                     DAPELFLKHGKGSVANDVTDEMVRLNWLTEFMPLPTIKHFIRTPDDAWLLTTAIPGKTA
                     FQVLEEYPDSGENIVDALAVFLRRLHSIPVCNCPFNSDRVFRLAQAQSRMNNGLVDASD
                     FDDERNGWPVEQVWKEMHKLLPFSPDSVVTHGDFSLDNLIFDEGKLIGCIDVGRVGIAD
                     RYQDLAILWNCLGEFSPSLQKRLFQKYGIDNPDMNKLQFHLMLDEFF"
     primer_bind     complement(4130..4149)
                     /label=Kan-R
                     /note="Kanamycin resistance gene, reverse primer"
ORIGIN
        1 ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa acaaaaaaac caccgctacc
       61 agcggtggtt tgtttgccgg atcaagagct accaactctt tttccgaagg taactggctt
      121 cagcagagcg cagataccaa atactgttct tctagtgtag ccgtagttag gccaccactt
      181 caagaactct gtagcaccgc ctacatacct cgctctgcta atcctgttac cagtggctgc
      241 tgccagtggc gataagtcgt gtcttaccgg gttggactca agacgatagt taccggataa
      301 ggcgcagcgg tcgggctgaa cggggggttc gtgcacacag cccagcttgg agcgaacgac
      361 ctacaccgaa ctgagatacc tacagcgtga gctatgagaa agcgccacgc ttcccgaagg
      421 gagaaaggcg gacaggtatc cggtaagcgg cagggtcgga acaggagagc gcacgaggga
      481 gcttccaggg ggaaacgcct ggtatcttta tagtcctgtc gggtttcgcc acctctgact
      541 tgagcgtcga tttttgtgat gctcgtcagg ggggcggagc ctatggaaaa acgccagcaa
      601 cgcggccttt ttacggttcc tggccttttg ctggcctttt gctcacatgt tctttcctgc
      661 gttatcccct gattctgtgg ataaccgtat taccgccttt gagtgagctg ataccgctcg
      721 ccgcagccga acgaccgagc gcagcgagtc agtgagcgag gaagcagagc gtgccacctg
      781 gtcgacttaa ccctagaaag ataatcatat tgtgacgtac gttaaagata atcatgcgta
      841 aaattgacgc atgtgtttta tcgatctgta tatcgaggtt tatttattaa tttgaataga
      901 tattaagttt tattatattt acacttacat actaataata aattcaacaa acaatttatt
      961 tatgtttatt tatttattaa aaaaaaacaa aaactcaaaa tttcttctat aaagtaacaa
     1021 aacttttaaa cattctctct tttacaaaaa taaacttatt ttgtacttta aaaacagtca
     1081 tgttgttctg caattcatgt gatctgcgcc tgggagctca cggggacagc ccccccccaa
     1141 agcccccagg gatgtaatta cgtccctccc ccgctagggg gcagcagcga gccgcccggg
     1201 gctccgctcc ggtccggcgc tccccccgca tccccgagcc ggcagcgtgc ggggacagcc
     1261 cgggcacggg gaaggtggca cgggatcgct ttcctctgaa cgcttctcgc tgctctttga
     1321 gcctgcagac acctgggggg atacggggaa aaagctaggc tgaaagagga tcttagaatg
     1381 acacgcgcct gggagctcac ggggacagcc cccccccaaa gcccccaggg atgtaattac
     1441 gtccctcccc cgctaggggg cagcagcgag ccgcccgggg ctccgctccg gtccggcgct
     1501 ccccccgcat ccccgagccg gcagcgtgcg gggacagccc gggcacgggg aaggtggcac
     1561 gggatcgctt tcctctgaac gcttctcgct gctctttgag cctgcagaca cctgggggga
     1621 tacggggaaa aagctggatc ataactcgac gtgcagtaaa acgacggcca ggcagtacca
     1681 gaagagcccg agtcttgcac tctgtggttt accgagctct tattggtttt caaacttcat
     1741 tgactgtgcc ttaattaata gggataacag ggtaatgtat cacgaggcag aatttcagat
     1801 aaaaaaaatc cttagctttc gctaaggatg atttctggaa ttcgcggccg cttctagagt
     1861 ccctatcagt gatagagatt gacatcccta tcagtgatag agatactgag cactactaga
     1921 gaaagaggag aaatactaga tggcttcctc cgaagacgtt atcaaagagt tcatgcgttt
     1981 caaagttcgt atggaaggtt ccgttaacgg tcacgagttc gaaatcgaag gtgaaggtga
     2041 aggtcgtccg tacgaaggta cccagaccgc taaactgaaa gttaccaaag gtggtccgct
     2101 gccgttcgct tgggacatcc tgtccccgca gttccagtac ggttccaaag cttacgttaa
     2161 acacccggct gacatcccgg actacctgaa actgtccttc ccggaaggtt tcaaatggga
     2221 acgtgttatg aacttcgaag acggtggtgt tgttaccgtt acccaggact cctccctgca
     2281 agacggtgag ttcatctaca aagttaaact gcgtggtacc aacttcccgt ccgacggtcc
     2341 ggttatgcag aaaaaaacca tgggttggga agcttccacc gaacgtatgt acccggaaga
     2401 cggtgctctg aaaggtgaaa tcaaaatgcg tctgaaactg aaagacggtg gtcactacga
     2461 cgctgaagtt aaaaccacct acatggctaa aaaaccggtt cagctgccgg gtgcttacaa
     2521 aaccgacatc aaactggaca tcacctccca caacgaagac tacaccatcg ttgaacagta
     2581 cgaacgtgct gaaggtcgtc actccaccgg tgcttaataa cgctgatagt gctagtgtag
     2641 atcgctacta gagccaggca tcaaataaaa cgaaaggctc agtcgaaaga ctgggccttt
     2701 cgttttatct gttgtttgtc ggtgaacgct ctctactaga gtcacactgg ctcaccttcg
     2761 ggtgggcctt tctgcgttta tatactagta ggatccatta ccctgttatc cctaaccggt
     2821 atcactagta ttacagaggt aagttataac agtcgcctaa ccacgaagtg ggctggacgc
     2881 gctcttcaag agttcgcttc tgcaattcat gtgatctgcg cctgggagct cacggggaca
     2941 gccccccccc aaagccccca gggatgtaat tacgtccctc ccccgctagg gggcagcagc
     3001 gagccgcccg gggctccgct ccggtccggc gctccccccg catccccgag ccggcagcgt
     3061 gcggggacag cccgggcacg gggaaggtgg cacgggatcg ctttcctctg aacgcttctc
     3121 gctgctcttt gagcctgcag acacctgggg ggatacgggg aaaaagctag gctgaaagag
     3181 gatcttagaa tgacacgcgc ctgggagctc acggggacag ccccccccca aagcccccag
     3241 ggatgtaatt acgtccctcc cccgctaggg ggcagcagcg agccgcccgg ggctccgctc
     3301 cggtccggcg ctccccccgc atccccgagc cggcagcgtg cggggacagc ccgggcacgg
     3361 ggaaggtggc acgggatcgc tttcctctga acgcttctcg ctgctctttg agcctgcaga
     3421 cacctggggg gatacgggga aaaagctgga tcataactcg acgtgcacag gctccagcct
     3481 acacaatcgc tcaagacgtg taatgctttt attatatatt agtcacgata tctataacaa
     3541 gaaaatatat atataataag ttatcacgta agtagaacat gaaataacaa tataattatc
     3601 gtatgagtta aatcttaaaa gtcacgtaaa agataatcat gcgtcatttt gactcacgcg
     3661 gtcgttatag ttcaaaatca gtgacactta ccgcattgac aagcacgcct cacgggagct
     3721 ccaagcggcg actgagatgt cctaaatgca cagcgacgga ttcgcgctat ttagaaagag
     3781 agagcaatat ttcaagaatg catgcgtcaa ttttacgcag actatctttc tagggttaaa
     3841 agcttggcgt aatcatgaat tcaggaaagg gcctcgtgat acgcctattt ttataggtta
     3901 atgtcatgat aataatggtt tcttagacgt caggtggcac ttttcgggga aatgtgctgg
     3961 cccgtgtctc aaaatctctg atgttacatt gcacaagata aaaatatatc atcatgaaca
     4021 ataaaactgt ctgcttacat aaacagtaat acaaggggtg ttatgagcca tattcaacgg
     4081 gaaacgtcga ggccgcgatt aaattccaac atggatgctg atttatatgg gtataaatgg
     4141 gctcgcgata atgtcgggca atcaggtgcg acaatctatc gcttgtatgg gaagcccgat
     4201 gcgccagagt tgtttctgaa acatggcaaa ggtagcgttg ccaatgatgt tacagatgag
     4261 atggtcagac taaactggct gacggaattt atgcctcttc cgaccatcaa gcattttatc
     4321 cgtactcctg atgatgcatg gttactcacc actgcgatcc ccggaaaaac agcattccag
     4381 gtattagaag aatatcctga ttcaggtgaa aatattgttg atgcgctggc agtgttcctg
     4441 cgccggttgc attcgattcc tgtttgtaat tgtcctttta acagcgatcg cgtatttcgt
     4501 ctcgctcagg cgcaatcacg aatgaataac ggtttggttg atgcgagtga ttttgatgac
     4561 gagcgtaatg gctggcctgt tgaacaagtc tggaaagaaa tgcataaact tttgccattc
     4621 tcaccggatt cagtcgtcac tcatggtgat ttctcacttg ataaccttat ttttgacgag
     4681 gggaaattaa taggttgtat tgatgttgga cgagtcggaa tcgcagaccg ataccaggat
     4741 cttgccatcc tatggaactg cctcggtgag ttttctcctt cattacagaa acggcttttt
     4801 caaaaatatg gtattgataa tcctgatatg aataaattgc agtttcattt gatgctcgat
     4861 gagtttttct aatcagaatt ggttaattgg ttgtaacact ggcagagcat tacgctgact
     4921 tgacgggacg gaagatcctt tttgataatc tcatgaccaa aatcccttaa cgtgagtttt
     4981 cgttccactg agcgtcagac cccgtagaaa agatcaaagg atcttc
//
