LOCUS       sequence-443610-        4897 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Donor plasmid of Tc1-8B_DR Transposon with copGFP mammalian 
            expression cassette.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 4897)
  AUTHORS   Zhang T, Tan S, Tang N, Li Y, Zhang C, Sun J, Guo Y, Gao H, Cai Y, 
            Sun W, Wang C, Fu L, Ma H, Wu Y, Hu X, Zhang X, Gee P, Yan W, Zhao 
            Y, Chen Q, Guo B, Wang H, Zhang YE
  TITLE     Heterologous survey of 130 DNA transposons in human cells highlights
            their functional divergence and expands the genome engineering 
            toolbox.
  JOURNAL   Cell. 2024 Jul 11;187(14):3741-3760.e30. doi: 
            10.1016/j.cell.2024.05.007. Epub 2024 Jun 5.
  PUBMED    38843831
REFERENCE   2  (bases 1 to 4897)
  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: 
            "10.1016/j.cell.2024.05.007"; journalName: "Cell"; date: 
            "2024-07-11- 11"; volume: "187"; issue: "14"; pages: "3741-3760.e30"
COMMENT     Sequence Label: pQLL-Tc1-8B_DR[EF1-alpha-copGFP]
FEATURES             Location/Qualifiers
     source          1..4897
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     complement(1151..1168)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21) 
                     Forward"
     primer_bind     complement(1151..1167)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1177..1195)
                     /label=pBRforEco
                     /note="pBR322 vectors, upsteam of EcoRI site, forward 
                     primer"
     terminator      1230..1259
                     /label=T3Te terminator
                     /note="phage T3 early transcription terminator"
     rep_origin      1281..1868
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1769..1788
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     terminator      complement(1880..1907)
                     /label=T7Te terminator
                     /note="phage T7 early transcription terminator"
     CDS             complement(1931..2122)
                     /codon_start=1
                     /gene="rop"
                     /product="Rop protein, which maintains plasmids at low copy
                     number"
                     /label=rop
                     /translation="MTKQEKTALNMARFIRSQTLTLLEKLNELDADEQADICESLHDHA
                     DELYRSCLARFGDDGENL"
     CDS             complement(join(2123..2914,2915..2983))
                     /codon_start=1
                     /gene="bla"
                     /product="beta-lactamase"
                     /label=AmpR
                     /note="confers resistance to ampicillin, carbenicillin, and
                     related antibiotics"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESLRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLATIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGSLLRSA
                     LPAGWFIADKSGAGERGSRGIIAELGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     primer_bind     2746..2765
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(2984..3086)
                     /label=cat promoter
                     /note="promoter of the E. coli cat gene encoding 
                     chloramphenicol acetyltransferase"
     terminator      complement(3087..3118)
                     /label=tonB terminator
                     /note="bidirectional E. coli tonB-P14 transcription 
                     terminator"
     primer_bind     3170..3187
                     /label=L4440
                     /note="L4440 vector, forward primer"
     primer_bind     3199..3215
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3199..3215
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3591..3672
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3628..3647)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3682..3701
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     CDS             complement(3715..4380)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     CDS             complement(3715..4380)
                     /codon_start=1
                     /product="green fluorescent protein 2 from Pontellina 
                     plumata, also known as ppluGFP2 (Shagin et al., 2004)"
                     /label=CopGFP
                     /translation="MPAMKIECRITGTLNGVEFELVGGGEGTPEQGRMTNKMKSTKGAL
                     TFSPYLLSHVMGYGFYHFGTYPSGYENPFLHAINNGGYTNTRIEKYEDGGVLHVSFSYR
                     YEAGRVIGDFKVVGTGFPEDSVIFTDKIIRSNATVEHLHPMGDNVLVGSFARTFSLRDG
                     GYYSFVVDSHMHFKSAIHPSILQNGGPMFAFRRVEELHSNTELGIVEYQHAFKTPIAFA
                     "
     promoter        complement(join(4422..4897,1..703))
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     intron          join(4431..4897,1..472)
                     /label=EF-1-alpha intron A
                     /note="intron upstream of the start codon of human 
                     EF-1-alpha"
     primer_bind     complement(4454..4474)
                     /label=EF1a-F
                     /note="Human elongation factor-1a promoter, forward primer"
ORIGIN
        1 ggcgagcttg agactacccc cgtccgattc tcggtggccg cgctcgcagg ccccgcctcg
       61 ccgaacatgt gcgctgggac gcacgggccc cgtcgccgcc cgcggcccca aaaaccgaaa
      121 taccagtgtg cagatcttgg cccgcattta caagactatc ttgccagaaa aaaagcgtcg
      181 cagcaggtca tcaaaaattt taaatggcta gagacttatc gaaagcagcg agacaggcgc
      241 gaaggtgcca ccagattcgc acgcggcggc cccagcgccc aggccaggcc tcaactcaag
      301 cacgaggcga aggggctcct taagcgcaag gcctcgaact ctcccaccca cttccaaccc
      361 gaagctcggg atcaagaatc acgtactgca gccaggtgga agtaattcaa ggcacgcaag
      421 ggccataacc cgtaaagagg ccaggcccgc gggaaccaca cacggcactt acctgtgttc
      481 tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac tactgcactt atatacggtt
      541 ctcccccacc ctcgggaaaa aggcggagcc agtacacgac atcactttcc cagtttaccc
      601 cgcgccacct tctctaggca ccgggtcaat tgccgacccc tccccccaac ttctcgggga
      661 ctgtgggcga tgtgcgctct gcccactgac gggcaccgga gccactagtc aataatcaat
      721 gcctctagca ccggtgaagt ctttcctgtg tgcctcctga gtaatgagaa gcctttatag
      781 gccatcaatt aggactaaag cagctgatat caattagtac tgatagggag cagggttttt
      841 ctctcaatac tgacagatct cctgactttc tatgccattt tgcaccttgt tatcttcatg
      901 tgttcaatac ttattccctg tgtcatttca ctttatttat tatgactcaa cttgtatact
      961 taaatgttct gatttctttg tatgaattca atatttggct tgatggctac atctggtagg
     1021 aattttgtgt cagtagccca cttagaaatc cccttactga taaaaatgct gatgtgtcaa
     1081 atacttattt tccccgctgt acatatgaga gacgcaagac actgcggatc gagacgcatt
     1141 gcgtggttcg actggccgtc gttttacaac gaaagggcct cgtgatacgc ctattgactc
     1201 cagcgtaact ggactgcaat caactcactg gctcaccttc acgggtgggc ctttcttcgg
     1261 tagaaaatca aaggatcttc ttgagatcct ttttttctgc gcgtaatctg ctgcttgcaa
     1321 acaaaaaaac caccgctacc agcggtggtt tgtttgccgg atcaagagct accaactctt
     1381 tttccgaggt aactggcttc agcagagcgc agataccaaa tactgttctt ctagtgtagc
     1441 cgtagttagg ccaccacttc aagaactctg tagcaccgcc tacatacctc gctctgctaa
     1501 tcctgttacc agtggctgct gccagtggcg ataagtcgtg tcttaccggg ttggactcaa
     1561 gacgatagtt accggataag gcgcagcggt cgggctgaac ggggggttcg tgcacacagc
     1621 ccagcttgga gcgaacgacc tacaccgaac tgagatacct acagcgtgag ctatgagaaa
     1681 gcgccacgct tcccgaaggg agaaaggcgg acaggtatcc ggtaagcggc agggtcggaa
     1741 caggagagcg cacgagggag cttccagggg gaaacgcctg gtatctttat agtcctgtcg
     1801 ggtttcgcca cctctgactt gagcatcgat ttttgtgatg ctcgtcaggg gggcggagcc
     1861 tatggaaaaa cgccagcaac gcagaaaggc ccacccgaag gtgagccagg tgattacatt
     1921 tgggccctca tcagaggttt tcaccgtcat caccgaaacg cgcgaggcag ctgcggtaaa
     1981 gctcatcagc gtggtcgtga agcgattcac agatgtctgc ctgttcatcc gcgtccagct
     2041 cgttgagttt ctccagaagc gttaatgtct ggcttctgat aaagcgggcc atgttaaggg
     2101 cggttttttc ctgtttggtc acttaccaat gcttaatcag tgaggcacct atctcagcga
     2161 tctgtctatt tcgttcatcc atagttgcct gactccccgt cgtgtagata actacgatgc
     2221 gggagggctt accatctggc cccagttctg caatgatacc gcgggaccca cgctcaccgg
     2281 ctccagattt atcagcaata aaccagccag ccgggagggc cgagcgcaga agtgatcctg
     2341 caactttatc cgcctccatc cagtctatta attgttgccg ggaagctaga gtaagtagtt
     2401 cgccagttaa tagtttgcgc aacgttgttg ccattgctac aggcatcgtg gtgtcacgct
     2461 cgtcgtttgg tatggcttca ttcagctccg gttcccaacg atcaaggcga gttacatgat
     2521 cccccatgtt gtgcaaaaaa gcggttagct ccttcggtcc tccgatcgtt gccagaagta
     2581 agttggccgc agtgttatca ctcatggtta tggcagcact gcataattct cttactgtca
     2641 tgccatccgt gagatgcttt tctgtgactg gtgagtactc aaccaagtca ttctgagaat
     2701 agtgtatgcg gcgaccgagt tgctcttgcc cggcgtcaat acgggataat accgcgccac
     2761 atagcagaac tttaaaagtg ctcatcattg gaaaacgttc ttcggggcgt aaactctcaa
     2821 ggatcttacc gctgttgaga tccagttcga tgtaacccac tcgtgcaccc aactgatctt
     2881 cagcatcttt tactttcacc agcgtttctg ggtgagcaaa aacaggaagg caaaatgccg
     2941 caaaaaaggg aataagggcg acacggaaat gttgaatact cattttagct tccttagctc
     3001 ctgaaaatct cgataactca aaaaatacgc ccggtagtga tcttatttca ttatggtgaa
     3061 agttggaacc tcttacgtgc cgatcaagtc aaaagcctcc ggtcggaggc ttttgacttt
     3121 ctgctatgga ggtcaggtat gatttaaatg gtcagtaacg accgagcgca gcgagtcagt
     3181 gagcgaggaa gcggcacaca ggaaacagct atgaccgtct cggttggcag tgactccgtc
     3241 tctgggccct acagcgggga aaataagtat ttgacacatc agcattttta tcagtaaggg
     3301 gatttctaag tgggctactg acacaaaatt cctaccagat gtagccatca agccaaatat
     3361 tgaattcata caaagaaatc agaacattta agtatacaag ttgagtcata ataaataaag
     3421 tgaaatgaca cagggaataa gtattgaaca cactttacct cgagggcgcg ccctcgccct
     3481 ttggtctccc cctcttaagt accacatttg tagaggtttt acttgcttta aaaaacctcc
     3541 cacacctccc cctgaacctg aaacataaaa tgaatgcaat tgttgttgtt aacttgttta
     3601 ttgcagctta taatggttac aaataaagca atagcatcac aaatttcaca aataaagcat
     3661 ttttttcact gcattctagt tgtggtttgt ccaaactcat cgagctcgag atctggcgaa
     3721 ggcgatgggg gtcttgaagg cgtgctggta ctccacgatg cccagctcgg tgttgctgtg
     3781 cagctcctcc acgcggcgga aggcgaacat ggggcccccg ttctgcagga tgctggggtg
     3841 gatggcgctc ttgaagtgca tgtggctgtc caccacgaag ctgtagtagc cgccgtcgcg
     3901 caggctgaag gtgcgggcga agctgcccac cagcacgtta tcgcccatgg ggtgcaggtg
     3961 ctccacggtg gcgttgctgc ggatgatctt gtcggtgaag atcacgctgt cctcggggaa
     4021 gccggtgccc accaccttga agtcgccgat cacgcggccg gcctcgtagc ggtagctgaa
     4081 gctcacgtgc agcacgccgc cgtcctcgta cttctcgatg cgggtgttgg tgtagccgcc
     4141 gttgttgatg gcgtgcagga aggggttctc gtagccgctg gggtaggtgc cgaagtggta
     4201 gaagccgtag cccatcacgt ggctcagcag gtaggggctg aaggtcaggg cgcctttggt
     4261 gctcttcatc ttgttggtca tgcggccctg ctcgggggtg ccctctccgc cgcccaccag
     4321 ctcgaactcc acgccgttca gggtgccggt gatgcggcac tcgatcttca tggcgggcat
     4381 ggtggcgacc ggtagcgcta gcggcttcgg taccggatcc ctcacgacac ctgaaatgga
     4441 agaaaaaaac tttgaaccac tgtctgaggc ttgagaatga accaagatcc aaactcaaaa
     4501 agggcaaatt ccaaggagaa ttacatcaag tgccaagctg gcctaacttc agtctccacc
     4561 cactcagtgt ggggaaactc catcgcataa aacccctccc cccaacctaa agacgacgta
     4621 ctccaaaagc tcgagaacta atcgaggtgc ctggacggcg cccggtactc agtggagtca
     4681 catgaagcga cggctgagga cggaaaggcc cttttccttt gtgtgggtga ctcacccgcc
     4741 cgctctcccg agcgccgcgt cctccatttt gagctccctg cagcagggcc gggaagcggc
     4801 catctttccg ctcacgcaac tggtgccgac cgggccagcc ttgccgccca gggcggggcg
     4861 atacacggcg gcgcgaggcc aggcaccaga gcaggcc
//
