LOCUS       sequence-442919-        5615 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of TC1_FR4 transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5615)
  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 5615)
  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: pCMV-TC1_FR4
FEATURES             Location/Qualifiers
     source          1..5615
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        complement(373..391)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(374..391)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(411..428)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    417..641
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      687..1115
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(774..793)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     984..1005
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(1124..1144)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        1129..1458
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      1309..1444
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     1371..1390
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    1624..1745
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(1661..1680)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     1715..1734
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(1794..1810)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(1794..1810)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(1807..1829)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    1818..1834
                     /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(1842..1848,1849..1866,1867..1872))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1887..1908
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(2025..2042)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(2196..2784)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(2276..2295)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(2955..3746,3747..3815))
                     /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     3578..3597
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(3816..3920)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(3995..4014)
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     enhancer        4186..4565
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     promoter        4566..4769
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     4720..4740
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     primer_bind     4814..4833
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        4814..4832
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     CDS             5573..5584
                     /codon_start=1
                     /product="Factor Xa recognition and cleavage site"
                     /label=Factor Xa site
                     /translation="IEGR"
ORIGIN
        1 aaaacaacgt gcaggagagc gtgaccaagc tgaaattacg tcgcggctgg gtttttcagc
       61 aggataacga tcctaaacat tgcagcaaaa gcacccgcgc ctttatgcag cgcaacaagt
      121 ataacgtgct ggagtggcct agccaaagcc ctgatctgaa catcatcgag aacctgtggg
      181 atgatctgaa acaggcggtg catgcacgca acccgagcaa cattaccgaa ctgcagcgct
      241 tttgccaagt ggagtgggcg aaaattccga gcagccgcat tcagaccctg attcaggcgg
      301 tgatttttgc gaaaggcggc agcaccaaat attaatgagc ggccgctcga gcatgcatct
      361 agagggccct attctatagt gtcacctaaa tgctagagct cgctgatcag cctcgactgt
      421 gccttctagt tgccagccat ctgttgtttg cccctccccc gtgccttcct tgaccctgga
      481 aggtgccact cccactgtcc tttcctaata aaatgaggaa attgcatcgc attgtctgag
      541 taggtgtcat tctattctgg ggggtggggt ggggcaggac agcaaggggg aggattggga
      601 agacaatagc aggcatgctg gggatgcggt gggctctatg gcttctgagg cggaaagaac
      661 cagctggggc tctagggggt atccccacgc gccctgtagc ggcgcattaa gcgcggcggg
      721 tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc ccgctccttt
      781 cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag ctctaaatcg
      841 ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca aaaaacttga
      901 ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc gccctttgac
      961 gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa cactcaaccc
     1021 tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct attggttaaa
     1081 aaatgagctg atttaacaaa aatttaacgc gaattaattc tgtggaatgt gtgtcagtta
     1141 gggtgtggaa agtccccagg ctccccagca ggcagaagta tgcaaagcat gcatctcaat
     1201 tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc
     1261 atgcatctca attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta
     1321 actccgccca gttccgccca ttctccgccc catggctgac taattttttt tatttatgca
     1381 gaggccgagg ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga
     1441 ggcctaggct tttgcaaaaa gctccccgaa atgaccgacc aagcgacgcc caacctgcca
     1501 tcacgagatt tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc
     1561 cgggacgccg gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac
     1621 cccaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc
     1681 acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta
     1741 tcttatcatg tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag
     1801 ctgtttcctg tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc
     1861 ataaagtgta aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc
     1921 tcactgcccg ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa
     1981 cgcgcgggga gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg
     2041 ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg
     2101 ttatccacag aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag
     2161 gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac
     2221 gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga
     2281 taccaggcgt ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt
     2341 accggatacc tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc
     2401 tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc
     2461 cccgttcagc ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta
     2521 agacacgact tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat
     2581 gtaggcggtg ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca
     2641 gtatttggta tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct
     2701 tgatccggca aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt
     2761 acgcgcagaa aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct
     2821 cagtggaacg aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc
     2881 acctagatcc ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa
     2941 acttggtctg acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta
     3001 tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc
     3061 ttaccatctg gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat
     3121 ttatcagcaa taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta
     3181 tccgcctcca tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt
     3241 aatagtttgc gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt
     3301 ggtatggctt cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg
     3361 ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc
     3421 gcagtgttat cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc
     3481 gtaagatgct tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg
     3541 cggcgaccga gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga
     3601 actttaaaag tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta
     3661 ccgctgttga gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct
     3721 tttactttca ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag
     3781 ggaataaggg cgacacggaa atgttgaata ctcatactct tcctttttca atattattga
     3841 agcatttatc agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat
     3901 aaacaaatag gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg
     3961 ggagatctcc cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt
     4021 taagccagta tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa
     4081 tttaagctac aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta
     4141 ggcgttttgc gctgcttcgc gatgtacggg ccagatatac gcgttgacat tgattattga
     4201 ctagttatta atagtaatca attacggggt cattagttca tagcccatat atggagttcc
     4261 gcgttacata acttacggta aatggcccgc ctggctgacc gcccaacgac ccccgcccat
     4321 tgacgtcaat aatgacgtat gttcccatag taacgccaat agggactttc cattgacgtc
     4381 aatgggtgga gtatttacgg taaactgccc acttggcagt acatcaagtg tatcatatgc
     4441 caagtacgcc ccctattgac gtcaatgacg gtaaatggcc cgcctggcat tatgcccagt
     4501 acatgacctt atgggacttt cctacttggc agtacatcta cgtattagtc atcgctatta
     4561 ccatggtgat gcggttttgg cagtacatca atgggcgtgg atagcggttt gactcacggg
     4621 gatttccaag tctccacccc attgacgtca atgggagttt gttttggcac caaaatcaac
     4681 gggactttcc aaaatgtcgt aacaactccg ccccattgac gcaaatgggc ggtaggcgtg
     4741 tacggtggga ggtctatata agcagagctc tctggctaac tagagaaccc actgcttact
     4801 ggcttatcga aattaatacg actcactata gggagaccca agcttggtac cgagctcgga
     4861 tccactagta acggccgcca gtgtgctgga attcgccgcc accatgcctg tgctgatcct
     4921 gtttgacagc ccgctgaaga gctgcaatat gggcgcgagc aaacagctga gcgattatct
     4981 gaagaccaag atcatccagc actatggcct gggcgaaggc tataaaaaac tgagcaagcg
     5041 cttccagctg agcgttagca ccgtgcgcaa cattgtgcgc aagtggaaag caaccggcac
     5101 cgttttagtg aaacctcgct cgggccgccc tcgtaaaatt gcggaacgtc agcgccgtcg
     5161 tatggttcgc atggtgaaaa acaaccctca gaccaccagc aaagatctgc agggctattt
     5221 agcggcagat ggtgtggcag ttcatcgtag caccatccag cacaacctgc acaaggaacg
     5281 cctgtacggt cgtgtgatgc gcaagaaacc gtttctgcat agccgccata agttaagccg
     5341 tctgcgctac gccaaagagc atctggataa accgattagc ttctggaaca aaatcctgtg
     5401 gaccgacgaa accaaaattg agctgtttgg ccacaacaaa cgccgttatg cgtggcgtca
     5461 gaaaaacacc gagttcgagg aaaaacatct gctgccgacc gttaaatttg gcggcggcag
     5521 cattatgtta tggggctgcg ttgcatcagc aggcactggt aacctggtta aaatcgaggg
     5581 ccgcatggat agcacccagt atcagcagat tctgg
//
