LOCUS       sequence-443030-        5645 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Tc1-5_Xt transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5645)
  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 5645)
  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-Tc1-5_Xt
FEATURES             Location/Qualifiers
     source          1..5645
                     /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"
     promoter        complement(1833..1851)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1834..1851)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1871..1888)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1877..2101
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2147..2575
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2234..2253)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2444..2465
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2584..2604)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2589..2918
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      2769..2904
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2831..2850
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3084..3205
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3121..3140)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3175..3194
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3254..3270)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3254..3270)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3267..3289)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3278..3294
                     /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(3302..3308,3309..3326,3327..3332))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3347..3368
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3485..3502)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3656..4244)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3736..3755)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4415..5206,5207..5275))
                     /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     5038..5057
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5276..5380)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5455..5474)
                     /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 gatcggctac aaaaaaagct ttagcgagtg gcaatgcctg agcgaagcga aaatgggtcg
      781 cggcagccct attcctacta tgctgcgccg caaaattgtg gagcagtacc agaagggtgt
      841 gacccagcgc aaaattgcga agatcctgca cttaagcagc agcaccgtgc acaacattat
      901 ccgccgcttt cgcgaaagcg gcaccatcag cgtgcgtaaa ggccaaggcc gcaaaaccat
      961 tttagacgcg cgcgatttac gtgcgctgaa gcgtcattgc accaccaatc gcaacgcgac
     1021 cgtgaaagag atcaccgaat gggcgcagga atatttccag aagccgctga gcgtgaacac
     1081 cattcatcgc gcgatccgtc gctgtcagct gaaactgtat agcgccaaga aaaagccgtt
     1141 tctgagcaaa atccataaac tgcgccgctt tcattgggca cgcgatcatc tgaaatggag
     1201 cgtggcgaaa tggaaaaccg tgctgtggag cgatgagagc cgctttgaag tgctgttcgg
     1261 caacttaggc cgccatgtga ttcgcaccaa ggaagataag gataacccga gctgctatca
     1321 gcgcagcgtg caaaaacctg cgagcctgat ggtttggggt tgcatgagcg cgtgtggcat
     1381 gggtagctta catgtgtgga agggcagcat taacgcggaa aagtatattc aggtgctgga
     1441 gcagcatatg ttacctagcc gccgccattt atttcaaggc cgcccgtgca tttttcagca
     1501 ggacaacgcc cgccctcata gcgcatcaat caccaccagc tggttacgtc gtcgtcgtat
     1561 tcgtgtgctg aaatggcctg tgtgcagccc tgatctgagc cctattgaaa acatctggcg
     1621 catcattaaa cgcaaagtgc gccagcgtcg ccctaaaacc atcgaacagc tggaagcgtg
     1681 catacgccag gaatgggaaa gcattccgat tccgaaactg gagaaactgg tgagcagcgt
     1741 gcctcgtcgt ttactgagcg tggttcgtcg tcgtggtgat gccacccaat ggtgatgagc
     1801 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     1861 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     1921 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     1981 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     2041 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     2101 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     2161 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     2221 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     2281 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     2341 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     2401 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     2461 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     2521 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     2581 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     2641 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     2701 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     2761 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     2821 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     2881 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgaa atgaccgacc
     2941 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     3001 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     3061 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     3121 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     3181 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     3241 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     3301 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     3361 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     3421 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     3481 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     3541 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     3601 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     3661 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     3721 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     3781 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     3841 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     3901 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     3961 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     4021 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     4081 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     4141 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     4201 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     4261 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     4321 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     4381 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     4441 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     4501 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     4561 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     4621 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     4681 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     4741 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     4801 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     4861 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     4921 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     4981 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     5041 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     5101 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     5161 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     5221 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     5281 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     5341 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     5401 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     5461 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     5521 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     5581 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     5641 gcgtt
//
