LOCUS       sequence-443175-        7205 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-4_Crp transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7205)
  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 7205)
  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-hAT-4_Crp
FEATURES             Location/Qualifiers
     source          1..7205
                     /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(3393..3411)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(3394..3411)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(3431..3448)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    3437..3661
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3707..4135
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3794..3813)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     4004..4025
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(4144..4164)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        4149..4478
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      4329..4464
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     4391..4410
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4644..4765
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4681..4700)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4735..4754
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4814..4830)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4814..4830)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4827..4849)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4838..4854
                     /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(4862..4868,4869..4886,4887..4892))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4907..4928
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(5045..5062)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(5216..5804)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(5296..5315)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5975..6766,6767..6835))
                     /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     6598..6617
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6836..6940)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(7015..7034)
                     /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 gagcggctac aaatataaaa gcggcagcca gaagcataaa gaggtgaaac agcgcgaact
      781 gaaagaagcg aaaggccatc agagcatcct gcagtttttt aaaaccgaca aggcctttct
      841 gcaaagcggc agcggcaata ccttaacccg cgaagtggat gagcagaaac aggacggcaa
      901 cgattttaac attgaaacgg tgagcgtggc caccgatgat gaagagaaac tgccggatgc
      961 cagctgtgac agcaacctga tgctgagcaa ggatagccag gtgattagcc cgctggcgcc
     1021 tttacgtagc gaagaaggcc tgcagaaaac cggtgaaagc agcaccaccc tgagcgatag
     1081 ctgctttttt aaagatcctg gcctgtggcc gaagctgatt accgatagcg tgcgccagaa
     1141 aattgtgctg agcggcgtgt taacctttga ggagatgtgg aaactggcga aaagcatccc
     1201 taaagatgtg ggcggccata agtttccgga gtttctgctg aagtgcaaaa ccagcaacgg
     1261 cgagcaactg cctcgtgatt ggttagtttg gagcccgacc aaccaagcgc tgtattgctt
     1321 tccgtgctgg ttactgtcag aaggccgcca aaaacagtgt agcatgctgg ccaaaagcga
     1381 aggctatagc ccgaacgtga aaaaatggca gaaactgtac aacgagctgc ctgagcatga
     1441 gaaaagcatt gaacacaaga cctgctattg ccagtggaag gaactgcagc agagcctgtc
     1501 aggtggtcat ggcgtggata gcgaagtgca gaaacagatt ctgaccgaag cgacccgctg
     1561 gaaagaaatt ctggagcgcc tgctggatgt gaccctgtat ttagcgagcc gcggtttagc
     1621 attccagggc gataacaacc tgattggcga tccgcataac ggcaactttc tgggcaccct
     1681 ggagttaatc gcccattacg atagcatcac ccgcgaccat ctggccaaag tgaaacagaa
     1741 gcagaccgag ggcaaaacca tgcaaggcca ggcccattat ctgagctggg aaagccagaa
     1801 cgaatttacc gaactgtgcg gcaaaagcgt gcagaaagcc attctggcgg agcgcgaaaa
     1861 agcgatttac tacagcatca tctgcgatgc caccccggat gtgtcacatc aggaacagca
     1921 tgtgctgatt ttccagttcg tggtgcataa caaggacacc ggcaccttta acattaaaga
     1981 gcgcttcttc gaatttatca acttcgacct gaaaaacggc gaacagatcg ccgagatgct
     2041 gattgcgagc ctgaaacgcc acaacattcc gctggatgat tgtcgtggcc agggctttga
     2101 taacggcagc aacatgagcg gcaagattaa aggcgtgcag gcctgcatta tgagcaaaaa
     2161 cgaactggcc gtgtatagcc cttgtgcgag ccatagctta aatctggtgg gcgtgaatgc
     2221 ggcagcggca tgtactgaaa ccctgacctt ttttggctgc gtgcagcagc tgtttgtgtt
     2281 tttcagcgcg agcccggtgc gctggaaaat tatcaccgac aagattcatt gcagcctgca
     2341 tagcctgagc gatacctgtt ggagcgcacg tgcggatgca attcgcccgt taaccaaaca
     2401 cctgccgggc attctggaag cactggaatg cgcgctggaa ctgaacctga cccataaggc
     2461 gcagagccaa ctggaagcgc tgaaacgcta ctttagcagc tttagctgct tactgctggc
     2521 gagcttttgg tataaagtgc tgagcagcat tgatgaccgc aacaaaattt tacaggcgcg
     2581 cggcattagc ttagaaagcg aggcagcgct gattgcggat attattgagg aagtgaagca
     2641 gctgcgtgac cagtggccta agattctgca ggaggcctgc ttagttgcag aaggcatggg
     2701 catactgcct cagctgccga aaaagcgcgg caagaaacag aaacgctttc acgatgacac
     2761 cgccgaagaa agctgcagca ccattagcga aaacgacccg gaaagcctgt ttcgccgcaa
     2821 tgtgttcgat gccaccctgg acaacgtgat ctatggtctg gagcgccgct ttaaagcgac
     2881 caacaacatc tgctttacct ttcgcagcgt gctgaaatat gtgaccctgg agcctgagga
     2941 actggataag gcgagcaaat gcctgctgca tcgctatcct accgacctga gcgaaaactt
     3001 tccgtgcgaa atgctgcatc tgaagaagaa cttcaaaagc accttcggca aagaatatga
     3061 actgccgccg ctgatcttac tgaacgagat ctacaaaaag cagctgcagc tgatctttag
     3121 cgaagtgtgc atcgcgattc gcattttttg caccctgcct gtgaccgttg cagaagcgga
     3181 acgcagcttt agcaagctgg cgctgattaa aaactgccgc cgcagcacta tgggtcaaga
     3241 gcgcttaagc catctggcga tcctgagcat cgagtgtgaa ctggcgcgcg agattgactt
     3301 tggcgaaatc atccacgact tcgcgatgaa aaaagcccgc aagatcaaac tgtgatgagc
     3361 ggccgctcga gcatgcatct agagggccct attctatagt gtcacctaaa tgctagagct
     3421 cgctgatcag cctcgactgt gccttctagt tgccagccat ctgttgtttg cccctccccc
     3481 gtgccttcct tgaccctgga aggtgccact cccactgtcc tttcctaata aaatgaggaa
     3541 attgcatcgc attgtctgag taggtgtcat tctattctgg ggggtggggt ggggcaggac
     3601 agcaaggggg aggattggga agacaatagc aggcatgctg gggatgcggt gggctctatg
     3661 gcttctgagg cggaaagaac cagctggggc tctagggggt atccccacgc gccctgtagc
     3721 ggcgcattaa gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc
     3781 gccctagcgc ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt
     3841 ccccgtcaag ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac
     3901 ctcgacccca aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag
     3961 acggtttttc gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa
     4021 actggaacaa cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg
     4081 atttcggcct attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattaattc
     4141 tgtggaatgt gtgtcagtta gggtgtggaa agtccccagg ctccccagca ggcagaagta
     4201 tgcaaagcat gcatctcaat tagtcagcaa ccaggtgtgg aaagtcccca ggctccccag
     4261 caggcagaag tatgcaaagc atgcatctca attagtcagc aaccatagtc ccgcccctaa
     4321 ctccgcccat cccgccccta actccgccca gttccgccca ttctccgccc catggctgac
     4381 taattttttt tatttatgca gaggccgagg ccgcctctgc ctctgagcta ttccagaagt
     4441 agtgaggagg cttttttgga ggcctaggct tttgcaaaaa gctccccgaa atgaccgacc
     4501 aagcgacgcc caacctgcca tcacgagatt tcgattccac cgccgccttc tatgaaaggt
     4561 tgggcttcgg aatcgttttc cgggacgccg gctggatgat cctccagcgc ggggatctca
     4621 tgctggagtt cttcgcccac cccaacttgt ttattgcagc ttataatggt tacaaataaa
     4681 gcaatagcat cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt
     4741 tgtccaaact catcaatgta tcttatcatg tctgtatacc gtcgacctct agctagagct
     4801 tggcgtaatc atggtcatag ctgtttcctg tgtgaaattg ttatccgctc acaattccac
     4861 acaacatacg agccggaagc ataaagtgta aagcctgggg tgcctaatga gtgagctaac
     4921 tcacattaat tgcgttgcgc tcactgcccg ctttccagtc gggaaacctg tcgtgccagc
     4981 tgcattaatg aatcggccaa cgcgcgggga gaggcggttt gcgtattggg cgctcttccg
     5041 cttcctcgct cactgactcg ctgcgctcgg tcgttcggct gcggcgagcg gtatcagctc
     5101 actcaaaggc ggtaatacgg ttatccacag aatcagggga taacgcagga aagaacatgt
     5161 gagcaaaagg ccagcaaaag gccaggaacc gtaaaaaggc cgcgttgctg gcgtttttcc
     5221 ataggctccg cccccctgac gagcatcaca aaaatcgacg ctcaagtcag aggtggcgaa
     5281 acccgacagg actataaaga taccaggcgt ttccccctgg aagctccctc gtgcgctctc
     5341 ctgttccgac cctgccgctt accggatacc tgtccgcctt tctcccttcg ggaagcgtgg
     5401 cgctttctca tagctcacgc tgtaggtatc tcagttcggt gtaggtcgtt cgctccaagc
     5461 tgggctgtgt gcacgaaccc cccgttcagc ccgaccgctg cgccttatcc ggtaactatc
     5521 gtcttgagtc caacccggta agacacgact tatcgccact ggcagcagcc actggtaaca
     5581 ggattagcag agcgaggtat gtaggcggtg ctacagagtt cttgaagtgg tggcctaact
     5641 acggctacac tagaagaaca gtatttggta tctgcgctct gctgaagcca gttaccttcg
     5701 gaaaaagagt tggtagctct tgatccggca aacaaaccac cgctggtagc ggtggttttt
     5761 ttgtttgcaa gcagcagatt acgcgcagaa aaaaaggatc tcaagaagat cctttgatct
     5821 tttctacggg gtctgacgct cagtggaacg aaaactcacg ttaagggatt ttggtcatga
     5881 gattatcaaa aaggatcttc acctagatcc ttttaaatta aaaatgaagt tttaaatcaa
     5941 tctaaagtat atatgagtaa acttggtctg acagttacca atgcttaatc agtgaggcac
     6001 ctatctcagc gatctgtcta tttcgttcat ccatagttgc ctgactcccc gtcgtgtaga
     6061 taactacgat acgggagggc ttaccatctg gccccagtgc tgcaatgata ccgcgagacc
     6121 cacgctcacc ggctccagat ttatcagcaa taaaccagcc agccggaagg gccgagcgca
     6181 gaagtggtcc tgcaacttta tccgcctcca tccagtctat taattgttgc cgggaagcta
     6241 gagtaagtag ttcgccagtt aatagtttgc gcaacgttgt tgccattgct acaggcatcg
     6301 tggtgtcacg ctcgtcgttt ggtatggctt cattcagctc cggttcccaa cgatcaaggc
     6361 gagttacatg atcccccatg ttgtgcaaaa aagcggttag ctccttcggt cctccgatcg
     6421 ttgtcagaag taagttggcc gcagtgttat cactcatggt tatggcagca ctgcataatt
     6481 ctcttactgt catgccatcc gtaagatgct tttctgtgac tggtgagtac tcaaccaagt
     6541 cattctgaga atagtgtatg cggcgaccga gttgctcttg cccggcgtca atacgggata
     6601 ataccgcgcc acatagcaga actttaaaag tgctcatcat tggaaaacgt tcttcggggc
     6661 gaaaactctc aaggatctta ccgctgttga gatccagttc gatgtaaccc actcgtgcac
     6721 ccaactgatc ttcagcatct tttactttca ccagcgtttc tgggtgagca aaaacaggaa
     6781 ggcaaaatgc cgcaaaaaag ggaataaggg cgacacggaa atgttgaata ctcatactct
     6841 tcctttttca atattattga agcatttatc agggttattg tctcatgagc ggatacatat
     6901 ttgaatgtat ttagaaaaat aaacaaatag gggttccgcg cacatttccc cgaaaagtgc
     6961 cacctgacgt cgacggatcg ggagatctcc cgatccccta tggtgcactc tcagtacaat
     7021 ctgctctgat gccgcatagt taagccagta tctgctccct gcttgtgtgt tggaggtcgc
     7081 tgagtagtgc gcgagcaaaa tttaagctac aacaaggcaa ggcttgaccg acaattgcat
     7141 gaagaatctg cttagggtta ggcgttttgc gctgcttcgc gatgtacggg ccagatatac
     7201 gcgtt
//
