LOCUS       sequence-442013-        5498 bp    DNA     circular SYN 15-APR-2025
DEFINITION  Mammalian expression vector for expression of Tc1-2_PM transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5498)
  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 5498)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 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: pCMV-Tc1-2_PM
FEATURES             Location/Qualifiers
     source          1..5498
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     enhancer        1..380
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        381..584
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     535..555
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     629..648
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        629..647
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     regulatory      713..722
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     RBS             844..852
                     /label=Shine-Dalgarno sequence
                     /note="full consensus sequence for ribosome-binding sites 
                     upstream of start codons in E. coli; complementary to a 
                     region in the 3' end of the 16S rRNA (Chen et al., 1994)"
     promoter        complement(1686..1704)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1687..1704)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1724..1741)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1730..1954
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2000..2428
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2087..2106)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2297..2318
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2437..2457)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2442..2771
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2622..2757
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2684..2703
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2937..3058
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2974..2993)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3028..3047
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3107..3123)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3107..3123)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3120..3142)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3131..3147
                     /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)."
     promoter        complement(join(3155..3161,3162..3179,3180..3185))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3200..3221
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3338..3355)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3509..4097)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3589..3608)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4268..5059,5060..5128))
                     /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     4891..4910
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5129..5233)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5308..5327)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
ORIGIN
        1 gacattgatt attgactagt tattaatagt aatcaattac ggggtcatta gttcatagcc
       61 catatatgga gttccgcgtt acataactta cggtaaatgg cccgcctggc tgaccgccca
      121 acgacccccg cccattgacg tcaataatga cgtatgttcc catagtaacg ccaataggga
      181 ctttccattg acgtcaatgg gtggagtatt tacggtaaac tgcccacttg gcagtacatc
      241 aagtgtatca tatgccaagt acgcccccta ttgacgtcaa tgacggtaaa tggcccgcct
      301 ggcattatgc ccagtacatg accttatggg actttcctac ttggcagtac atctacgtat
      361 tagtcatcgc tattaccatg gtgatgcggt tttggcagta catcaatggg cgtggatagc
      421 ggtttgactc acggggattt ccaagtctcc accccattga cgtcaatggg agtttgtttt
      481 ggcaccaaaa tcaacgggac tttccaaaat gtcgtaacaa ctccgcccca ttgacgcaaa
      541 tgggcggtag gcgtgtacgg tgggaggtct atataagcag agctctctgg ctaactagag
      601 aacccactgc ttactggctt atcgaaatta atacgactca ctatagggag acccaagctt
      661 ggtaccgagc tcggatccac tagtaacggc cgccagtgtg ctggaattcg ccgccaccat
      721 gggcttacat cagagcaccg tgcgccagat cgtgtacaaa tggcgcaaat tcaaaaccac
      781 cgtgacctta cctcgttcag gccgtcctac caaaataagc cctcgcacca accgcaaaat
      841 tattaaggag gtgaccaaaa accctcgcga aaccagcaaa gatctgcagg cgaccttagc
      901 actggcgaat gtgagcgtgc acgacagcac cattcgcaaa cgcctgaaca aaaacggtgt
      961 gcacggtcgt atagcacgcc gcaaaccttt actgagcaag aagaacattg cggcgcgcct
     1021 gaaatttgcg aaagagcaca ttgatgaccc tcaggatttc tggaacaacg tgctgtggac
     1081 cgacgaaagc aaggtggaac tgtttggcct gaacgagaaa cgctatgtgt ggcgcaaacc
     1141 gaacaccgcg tttgaacaga aaaacctgat ccctaccgtt aaacgcggcg gtggtaacgt
     1201 tatggtttgg ggctgctttg cagcgagcgg tcctggtcgt ctggccatta ttgataccac
     1261 catgaacagc gccctgtatc agaagattct ggaagagaac gtgcgtcctt cagtgcgcga
     1321 actgaaactg aaccgcaaat gggtgctgca gcaggataac gatccgaaac acaccagccg
     1381 cagcaccaaa gaatggctgc agaagaaaaa gtttcgcgtg ttacagtggc ctagccaaag
     1441 ccctgatctg aacccgattg agatgctgtg gcaggatctg aaacgtgcgg tgcacgcacg
     1501 tcagcctagc aatgttaccg agctgaaaga gttttgcaaa gaagagtggg cgaagatccc
     1561 tcagattcgc tgcgaacgtc tgactaacgg ctatcgcaaa cgcctggtgg cagttatagc
     1621 agcccaaggt ggtgttacca gctattgatg agcggccgct cgagcatgca tctagagggc
     1681 cctattctat agtgtcacct aaatgctaga gctcgctgat cagcctcgac tgtgccttct
     1741 agttgccagc catctgttgt ttgcccctcc cccgtgcctt ccttgaccct ggaaggtgcc
     1801 actcccactg tcctttccta ataaaatgag gaaattgcat cgcattgtct gagtaggtgt
     1861 cattctattc tggggggtgg ggtggggcag gacagcaagg gggaggattg ggaagacaat
     1921 agcaggcatg ctggggatgc ggtgggctct atggcttctg aggcggaaag aaccagctgg
     1981 ggctctaggg ggtatcccca cgcgccctgt agcggcgcat taagcgcggc gggtgtggtg
     2041 gttacgcgca gcgtgaccgc tacacttgcc agcgccctag cgcccgctcc tttcgctttc
     2101 ttcccttcct ttctcgccac gttcgccggc tttccccgtc aagctctaaa tcgggggctc
     2161 cctttagggt tccgatttag tgctttacgg cacctcgacc ccaaaaaact tgattagggt
     2221 gatggttcac gtagtgggcc atcgccctga tagacggttt ttcgcccttt gacgttggag
     2281 tccacgttct ttaatagtgg actcttgttc caaactggaa caacactcaa ccctatctcg
     2341 gtctattctt ttgatttata agggattttg ccgatttcgg cctattggtt aaaaaatgag
     2401 ctgatttaac aaaaatttaa cgcgaattaa ttctgtggaa tgtgtgtcag ttagggtgtg
     2461 gaaagtcccc aggctcccca gcaggcagaa gtatgcaaag catgcatctc aattagtcag
     2521 caaccaggtg tggaaagtcc ccaggctccc cagcaggcag aagtatgcaa agcatgcatc
     2581 tcaattagtc agcaaccata gtcccgcccc taactccgcc catcccgccc ctaactccgc
     2641 ccagttccgc ccattctccg ccccatggct gactaatttt ttttatttat gcagaggccg
     2701 aggccgcctc tgcctctgag ctattccaga agtagtgagg aggctttttt ggaggcctag
     2761 gcttttgcaa aaagctcccc gaaatgaccg accaagcgac gcccaacctg ccatcacgag
     2821 atttcgattc caccgccgcc ttctatgaaa ggttgggctt cggaatcgtt ttccgggacg
     2881 ccggctggat gatcctccag cgcggggatc tcatgctgga gttcttcgcc caccccaact
     2941 tgtttattgc agcttataat ggttacaaat aaagcaatag catcacaaat ttcacaaata
     3001 aagcattttt ttcactgcat tctagttgtg gtttgtccaa actcatcaat gtatcttatc
     3061 atgtctgtat accgtcgacc tctagctaga gcttggcgta atcatggtca tagctgtttc
     3121 ctgtgtgaaa ttgttatccg ctcacaattc cacacaacat acgagccgga agcataaagt
     3181 gtaaagcctg gggtgcctaa tgagtgagct aactcacatt aattgcgttg cgctcactgc
     3241 ccgctttcca gtcgggaaac ctgtcgtgcc agctgcatta atgaatcggc caacgcgcgg
     3301 ggagaggcgg tttgcgtatt gggcgctctt ccgcttcctc gctcactgac tcgctgcgct
     3361 cggtcgttcg gctgcggcga gcggtatcag ctcactcaaa ggcggtaata cggttatcca
     3421 cagaatcagg ggataacgca ggaaagaaca tgtgagcaaa aggccagcaa aaggccagga
     3481 accgtaaaaa ggccgcgttg ctggcgtttt tccataggct ccgcccccct gacgagcatc
     3541 acaaaaatcg acgctcaagt cagaggtggc gaaacccgac aggactataa agataccagg
     3601 cgtttccccc tggaagctcc ctcgtgcgct ctcctgttcc gaccctgccg cttaccggat
     3661 acctgtccgc ctttctccct tcgggaagcg tggcgctttc tcatagctca cgctgtaggt
     3721 atctcagttc ggtgtaggtc gttcgctcca agctgggctg tgtgcacgaa ccccccgttc
     3781 agcccgaccg ctgcgcctta tccggtaact atcgtcttga gtccaacccg gtaagacacg
     3841 acttatcgcc actggcagca gccactggta acaggattag cagagcgagg tatgtaggcg
     3901 gtgctacaga gttcttgaag tggtggccta actacggcta cactagaaga acagtatttg
     3961 gtatctgcgc tctgctgaag ccagttacct tcggaaaaag agttggtagc tcttgatccg
     4021 gcaaacaaac caccgctggt agcggtggtt tttttgtttg caagcagcag attacgcgca
     4081 gaaaaaaagg atctcaagaa gatcctttga tcttttctac ggggtctgac gctcagtgga
     4141 acgaaaactc acgttaaggg attttggtca tgagattatc aaaaaggatc ttcacctaga
     4201 tccttttaaa ttaaaaatga agttttaaat caatctaaag tatatatgag taaacttggt
     4261 ctgacagtta ccaatgctta atcagtgagg cacctatctc agcgatctgt ctatttcgtt
     4321 catccatagt tgcctgactc cccgtcgtgt agataactac gatacgggag ggcttaccat
     4381 ctggccccag tgctgcaatg ataccgcgag acccacgctc accggctcca gatttatcag
     4441 caataaacca gccagccgga agggccgagc gcagaagtgg tcctgcaact ttatccgcct
     4501 ccatccagtc tattaattgt tgccgggaag ctagagtaag tagttcgcca gttaatagtt
     4561 tgcgcaacgt tgttgccatt gctacaggca tcgtggtgtc acgctcgtcg tttggtatgg
     4621 cttcattcag ctccggttcc caacgatcaa ggcgagttac atgatccccc atgttgtgca
     4681 aaaaagcggt tagctccttc ggtcctccga tcgttgtcag aagtaagttg gccgcagtgt
     4741 tatcactcat ggttatggca gcactgcata attctcttac tgtcatgcca tccgtaagat
     4801 gcttttctgt gactggtgag tactcaacca agtcattctg agaatagtgt atgcggcgac
     4861 cgagttgctc ttgcccggcg tcaatacggg ataataccgc gccacatagc agaactttaa
     4921 aagtgctcat cattggaaaa cgttcttcgg ggcgaaaact ctcaaggatc ttaccgctgt
     4981 tgagatccag ttcgatgtaa cccactcgtg cacccaactg atcttcagca tcttttactt
     5041 tcaccagcgt ttctgggtga gcaaaaacag gaaggcaaaa tgccgcaaaa aagggaataa
     5101 gggcgacacg gaaatgttga atactcatac tcttcctttt tcaatattat tgaagcattt
     5161 atcagggtta ttgtctcatg agcggataca tatttgaatg tatttagaaa aataaacaaa
     5221 taggggttcc gcgcacattt ccccgaaaag tgccacctga cgtcgacgga tcgggagatc
     5281 tcccgatccc ctatggtgca ctctcagtac aatctgctct gatgccgcat agttaagcca
     5341 gtatctgctc cctgcttgtg tgttggaggt cgctgagtag tgcgcgagca aaatttaagc
     5401 tacaacaagg caaggcttga ccgacaattg catgaagaat ctgcttaggg ttaggcgttt
     5461 tgcgctgctt cgcgatgtac gggccagata tacgcgtt
//
