LOCUS       sequence-443035-        5495 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of Tc1-1_AG transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5495)
  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 5495)
  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-Tc1-1_AG
FEATURES             Location/Qualifiers
     source          1..5495
                     /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"
     promoter        complement(1755..1773)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(1756..1773)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(1793..1810)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    1799..2023
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2069..2497
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2156..2175)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2366..2387
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2506..2526)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2550..2760
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        2572..2768
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     rep_origin      2619..2754
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     2681..2700
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    2934..3055
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(2971..2990)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3025..3044
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3104..3120)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3104..3120)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3117..3139)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3128..3144
                     /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(3152..3158,3159..3176,3177..3182))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3197..3218
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3335..3352)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(3506..4094)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(3586..3605)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4265..5056,5057..5125))
                     /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     4888..4907
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5126..5230)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5305..5324)
                     /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 gggtcgtggt aagcattgca ctcctgaaga gcgcaagcat attcagggcc tgtatcgcga
      781 aaacgtgccg atcaagacca tttgcaaggc gttcggccgt agccgtacct ttgtggataa
      841 cgccattcgc agcgaagcca ccggtaaaag caccggccgt ccgcgtaaaa ctaccgcaga
      901 tgtggatgcg cagattgtgg agatgattcg cgcggacccg tttaaaacct gcacccgcat
      961 taaacaggag ctgggcctgc aagttagcgc caagaccgtt agccgtcgtt tacatgccgc
     1021 aggtttttgt gcacgtcgtc cgcgtaaggt tcgtaaactg ctgccgcatc acgttgaagc
     1081 gcgcattcgc tttgccgaag aacatctggc ggcgagcatt ttttggtgga gcaagattat
     1141 cttcagcgac gaaagccgca ttaacctgga tggcagcgat ggcattaaat atgtgtggcg
     1201 cttccctaac caggcgtatc atccgaaaaa caccattaag accctgagcc atggtggtgg
     1261 tcacgtgatg gtttggggct gctttagctg gcatggcacc ggtcctctgt ttcgcattaa
     1321 cggcaccctg aacagcgaag gctatcgcaa gattctgagc cgcaaaatgc tgccttacgc
     1381 gcgtcagcag tttggcgatg aggagcatta catttttcag cacgacaacg atagcaaaca
     1441 caccagccgc accgtgaaat gctatctggc gaaccaggat gtgcaggtgt taccttggcc
     1501 tgcgttaagc ccggatctga accctatcga aaacctgtgg agcaccttaa agcgccagct
     1561 gaaaaatcaa cctgcgcgca gcgcagatga tttatggacc cgctgcaagg ttatgtggga
     1621 acgcatcccg cgtagcgaat gccgtaacct gattggcgat atggcgaaac gctgccagga
     1681 agtgattgcc aacaacggcc accagattga tcgctaatga gcggccgctc gagcatgcat
     1741 ctagagggcc ctattctata gtgtcaccta aatgctagag ctcgctgatc agcctcgact
     1801 gtgccttcta gttgccagcc atctgttgtt tgcccctccc ccgtgccttc cttgaccctg
     1861 gaaggtgcca ctcccactgt cctttcctaa taaaatgagg aaattgcatc gcattgtctg
     1921 agtaggtgtc attctattct ggggggtggg gtggggcagg acagcaaggg ggaggattgg
     1981 gaagacaata gcaggcatgc tggggatgcg gtgggctcta tggcttctga ggcggaaaga
     2041 accagctggg gctctagggg gtatccccac gcgccctgta gcggcgcatt aagcgcggcg
     2101 ggtgtggtgg ttacgcgcag cgtgaccgct acacttgcca gcgccctagc gcccgctcct
     2161 ttcgctttct tcccttcctt tctcgccacg ttcgccggct ttccccgtca agctctaaat
     2221 cgggggctcc ctttagggtt ccgatttagt gctttacggc acctcgaccc caaaaaactt
     2281 gattagggtg atggttcacg tagtgggcca tcgccctgat agacggtttt tcgccctttg
     2341 acgttggagt ccacgttctt taatagtgga ctcttgttcc aaactggaac aacactcaac
     2401 cctatctcgg tctattcttt tgatttataa gggattttgc cgatttcggc ctattggtta
     2461 aaaaatgagc tgatttaaca aaaatttaac gcgaattaat tctgtggaat gtgtgtcagt
     2521 tagggtgtgg aaagtcccca ggctccccag caggcagaag tatgcaaagc atgcatctca
     2581 attagtcagc aaccatagtc ccgcccctaa ctccgcccat cccgccccta actccgccca
     2641 gttccgccca ttctccgccc catggctgac taattttttt tatttatgca gaggccgagg
     2701 ccgcctctgc ctctgagcta ttccagaagt agtgaggagg cttttttgga ggcctaggct
     2761 tttgcaaaaa gctccccgaa atgaccgacc aagcgacgcc caacctgcca tcacgagatt
     2821 tcgattccac cgccgccttc tatgaaaggt tgggcttcgg aatcgttttc cgggacgccg
     2881 gctggatgat cctccagcgc ggggatctca tgctggagtt cttcgcccac cccaacttgt
     2941 ttattgcagc ttataatggt tacaaataaa gcaatagcat cacaaatttc acaaataaag
     3001 catttttttc actgcattct agttgtggtt tgtccaaact catcaatgta tcttatcatg
     3061 tctgtatacc gtcgacctct agctagagct tggcgtaatc atggtcatag ctgtttcctg
     3121 tgtgaaattg ttatccgctc acaattccac acaacatacg agccggaagc ataaagtgta
     3181 aagcctgggg tgcctaatga gtgagctaac tcacattaat tgcgttgcgc tcactgcccg
     3241 ctttccagtc gggaaacctg tcgtgccagc tgcattaatg aatcggccaa cgcgcgggga
     3301 gaggcggttt gcgtattggg cgctcttccg cttcctcgct cactgactcg ctgcgctcgg
     3361 tcgttcggct gcggcgagcg gtatcagctc actcaaaggc ggtaatacgg ttatccacag
     3421 aatcagggga taacgcagga aagaacatgt gagcaaaagg ccagcaaaag gccaggaacc
     3481 gtaaaaaggc cgcgttgctg gcgtttttcc ataggctccg cccccctgac gagcatcaca
     3541 aaaatcgacg ctcaagtcag aggtggcgaa acccgacagg actataaaga taccaggcgt
     3601 ttccccctgg aagctccctc gtgcgctctc ctgttccgac cctgccgctt accggatacc
     3661 tgtccgcctt tctcccttcg ggaagcgtgg cgctttctca tagctcacgc tgtaggtatc
     3721 tcagttcggt gtaggtcgtt cgctccaagc tgggctgtgt gcacgaaccc cccgttcagc
     3781 ccgaccgctg cgccttatcc ggtaactatc gtcttgagtc caacccggta agacacgact
     3841 tatcgccact ggcagcagcc actggtaaca ggattagcag agcgaggtat gtaggcggtg
     3901 ctacagagtt cttgaagtgg tggcctaact acggctacac tagaagaaca gtatttggta
     3961 tctgcgctct gctgaagcca gttaccttcg gaaaaagagt tggtagctct tgatccggca
     4021 aacaaaccac cgctggtagc ggtggttttt ttgtttgcaa gcagcagatt acgcgcagaa
     4081 aaaaaggatc tcaagaagat cctttgatct tttctacggg gtctgacgct cagtggaacg
     4141 aaaactcacg ttaagggatt ttggtcatga gattatcaaa aaggatcttc acctagatcc
     4201 ttttaaatta aaaatgaagt tttaaatcaa tctaaagtat atatgagtaa acttggtctg
     4261 acagttacca atgcttaatc agtgaggcac ctatctcagc gatctgtcta tttcgttcat
     4321 ccatagttgc ctgactcccc gtcgtgtaga taactacgat acgggagggc ttaccatctg
     4381 gccccagtgc tgcaatgata ccgcgagacc cacgctcacc ggctccagat ttatcagcaa
     4441 taaaccagcc agccggaagg gccgagcgca gaagtggtcc tgcaacttta tccgcctcca
     4501 tccagtctat taattgttgc cgggaagcta gagtaagtag ttcgccagtt aatagtttgc
     4561 gcaacgttgt tgccattgct acaggcatcg tggtgtcacg ctcgtcgttt ggtatggctt
     4621 cattcagctc cggttcccaa cgatcaaggc gagttacatg atcccccatg ttgtgcaaaa
     4681 aagcggttag ctccttcggt cctccgatcg ttgtcagaag taagttggcc gcagtgttat
     4741 cactcatggt tatggcagca ctgcataatt ctcttactgt catgccatcc gtaagatgct
     4801 tttctgtgac tggtgagtac tcaaccaagt cattctgaga atagtgtatg cggcgaccga
     4861 gttgctcttg cccggcgtca atacgggata ataccgcgcc acatagcaga actttaaaag
     4921 tgctcatcat tggaaaacgt tcttcggggc gaaaactctc aaggatctta ccgctgttga
     4981 gatccagttc gatgtaaccc actcgtgcac ccaactgatc ttcagcatct tttactttca
     5041 ccagcgtttc tgggtgagca aaaacaggaa ggcaaaatgc cgcaaaaaag ggaataaggg
     5101 cgacacggaa atgttgaata ctcatactct tcctttttca atattattga agcatttatc
     5161 agggttattg tctcatgagc ggatacatat ttgaatgtat ttagaaaaat aaacaaatag
     5221 gggttccgcg cacatttccc cgaaaagtgc cacctgacgt cgacggatcg ggagatctcc
     5281 cgatccccta tggtgcactc tcagtacaat ctgctctgat gccgcatagt taagccagta
     5341 tctgctccct gcttgtgtgt tggaggtcgc tgagtagtgc gcgagcaaaa tttaagctac
     5401 aacaaggcaa ggcttgaccg acaattgcat gaagaatctg cttagggtta ggcgttttgc
     5461 gctgcttcgc gatgtacggg ccagatatac gcgtt
//
