LOCUS       sequence-443173-        6020 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-3_Gav transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6020)
  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 6020)
  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-3_Gav
FEATURES             Location/Qualifiers
     source          1..6020
                     /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(2208..2226)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2209..2226)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2246..2263)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2252..2476
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2522..2950
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2609..2628)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     2819..2840
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(2959..2979)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        2964..3293
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3144..3279
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3206..3225
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3459..3580
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3496..3515)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3550..3569
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(3629..3645)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(3629..3645)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(3642..3664)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    3653..3669
                     /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(3677..3683,3684..3701,3702..3707))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3722..3743
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(3860..3877)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4031..4619)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4111..4130)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(4790..5581,5582..5650))
                     /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     5413..5432
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(5651..5755)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(5830..5849)
                     /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 ggacggtttt gtggtgcgca ccagcaagta tcagcaggac accatcgatg aaaaaatcgc
      781 gcgctttatt tatgcgacca acagcccgtt tcgcattgtg aacaacaaac actttatcga
      841 catggtggaa agcttacgcc cgggctatac tcctcctggt cgcgcagata taggtggtcg
      901 tctgctggat accgcctacg aaaaagaact ggaacagtgc gcgcgtaata ccgaaggcaa
      961 gtttgccagc ttaagcctgg atggctggag caacgtgcat aacgaccctg tgatatgcgc
     1021 ctgtgtgacc actgaagatg gcgtggcgta tctgagcgaa accatcgata ccagcggcaa
     1081 tgcccatacc gccgaatacc tgaaagaagt ggcggtgaaa gcggtgatta actgcgaaca
     1141 gaagtttaac tgccatgtgc gcagcttcgt gaccgataac gcagcgaacg tggcaaaaat
     1201 gcgccgctat ctggaagaag aaacggaagg ctgcagcctg attacctatg gctgcagcgc
     1261 gcacttaatg catctgctgg cgaaggattt atcaaccagc ccgggcatta aagaaaacgt
     1321 ggtggaggtg gcgaaatact tccgcaacaa ccattttgcc agcgcgagct taaaacgtgc
     1381 gggcggcagc aaattaatac tgccgcagga tgtgcgttgg aacagcgtgg tggattgctt
     1441 cgaacagttc attaagaact ggcccatcct gatcaaaatc tgcgaagaga accgcgataa
     1501 aattgatggc agcattagca gcaaagtgag caacatcggc ctgaagcgca acgtggaaga
     1561 tatgctgagc attctgaaac ctatcagcat tgcgctgaac aaactgcaac gcgactgctg
     1621 ttgtattgcg gacgcggtgg aaatttggaa ggcgctgcag gaaaccctga aaaaagaaat
     1681 cccgaaccag aaagtgaaac tgcaggcgat caagaaacgc atggaccagg cgttaacccc
     1741 gcctcatttt ctggccaaca tcctgaaccc gcgctataag ggccgttgtc tgaccagcga
     1801 agaagatgcg gcagcaatgg cgtgggcgag ccaaaatcat agcagcgtga tgccggtgat
     1861 tattaacttt cgcgccggcg gcgaaccttt taagcagtac atgtttaccg acgatgtgct
     1921 gaacaaggtg acccctctga actggtggaa aagcctggcg cgccacttag aaaccgagtt
     1981 cctgcagtta ctgagccagc tgtttaccgc agttgcgagc agcgcaggta ccgaacgcat
     2041 ctttagcagc ttcggcatta tccatagcaa gctgcgtaac cgtctgggcg ttgaaaaagc
     2101 gggcaagctg gtgttcctgt ttcagagcat gaacaaaaac gatagcggca gcggcgaaga
     2161 cgagatttga tgagcggccg ctcgagcatg catctagagg gccctattct atagtgtcac
     2221 ctaaatgcta gagctcgctg atcagcctcg actgtgcctt ctagttgcca gccatctgtt
     2281 gtttgcccct cccccgtgcc ttccttgacc ctggaaggtg ccactcccac tgtcctttcc
     2341 taataaaatg aggaaattgc atcgcattgt ctgagtaggt gtcattctat tctggggggt
     2401 ggggtggggc aggacagcaa gggggaggat tgggaagaca atagcaggca tgctggggat
     2461 gcggtgggct ctatggcttc tgaggcggaa agaaccagct ggggctctag ggggtatccc
     2521 cacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg cagcgtgacc
     2581 gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc ctttctcgcc
     2641 acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg gttccgattt
     2701 agtgctttac ggcacctcga ccccaaaaaa cttgattagg gtgatggttc acgtagtggg
     2761 ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt ctttaatagt
     2821 ggactcttgt tccaaactgg aacaacactc aaccctatct cggtctattc ttttgattta
     2881 taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta acaaaaattt
     2941 aacgcgaatt aattctgtgg aatgtgtgtc agttagggtg tggaaagtcc ccaggctccc
     3001 cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccagg tgtggaaagt
     3061 ccccaggctc cccagcaggc agaagtatgc aaagcatgca tctcaattag tcagcaacca
     3121 tagtcccgcc cctaactccg cccatcccgc ccctaactcc gcccagttcc gcccattctc
     3181 cgccccatgg ctgactaatt ttttttattt atgcagaggc cgaggccgcc tctgcctctg
     3241 agctattcca gaagtagtga ggaggctttt ttggaggcct aggcttttgc aaaaagctcc
     3301 ccgaaatgac cgaccaagcg acgcccaacc tgccatcacg agatttcgat tccaccgccg
     3361 ccttctatga aaggttgggc ttcggaatcg ttttccggga cgccggctgg atgatcctcc
     3421 agcgcgggga tctcatgctg gagttcttcg cccaccccaa cttgtttatt gcagcttata
     3481 atggttacaa ataaagcaat agcatcacaa atttcacaaa taaagcattt ttttcactgc
     3541 attctagttg tggtttgtcc aaactcatca atgtatctta tcatgtctgt ataccgtcga
     3601 cctctagcta gagcttggcg taatcatggt catagctgtt tcctgtgtga aattgttatc
     3661 cgctcacaat tccacacaac atacgagccg gaagcataaa gtgtaaagcc tggggtgcct
     3721 aatgagtgag ctaactcaca ttaattgcgt tgcgctcact gcccgctttc cagtcgggaa
     3781 acctgtcgtg ccagctgcat taatgaatcg gccaacgcgc ggggagaggc ggtttgcgta
     3841 ttgggcgctc ttccgcttcc tcgctcactg actcgctgcg ctcggtcgtt cggctgcggc
     3901 gagcggtatc agctcactca aaggcggtaa tacggttatc cacagaatca ggggataacg
     3961 caggaaagaa catgtgagca aaaggccagc aaaaggccag gaaccgtaaa aaggccgcgt
     4021 tgctggcgtt tttccatagg ctccgccccc ctgacgagca tcacaaaaat cgacgctcaa
     4081 gtcagaggtg gcgaaacccg acaggactat aaagatacca ggcgtttccc cctggaagct
     4141 ccctcgtgcg ctctcctgtt ccgaccctgc cgcttaccgg atacctgtcc gcctttctcc
     4201 cttcgggaag cgtggcgctt tctcatagct cacgctgtag gtatctcagt tcggtgtagg
     4261 tcgttcgctc caagctgggc tgtgtgcacg aaccccccgt tcagcccgac cgctgcgcct
     4321 tatccggtaa ctatcgtctt gagtccaacc cggtaagaca cgacttatcg ccactggcag
     4381 cagccactgg taacaggatt agcagagcga ggtatgtagg cggtgctaca gagttcttga
     4441 agtggtggcc taactacggc tacactagaa gaacagtatt tggtatctgc gctctgctga
     4501 agccagttac cttcggaaaa agagttggta gctcttgatc cggcaaacaa accaccgctg
     4561 gtagcggtgg tttttttgtt tgcaagcagc agattacgcg cagaaaaaaa ggatctcaag
     4621 aagatccttt gatcttttct acggggtctg acgctcagtg gaacgaaaac tcacgttaag
     4681 ggattttggt catgagatta tcaaaaagga tcttcaccta gatcctttta aattaaaaat
     4741 gaagttttaa atcaatctaa agtatatatg agtaaacttg gtctgacagt taccaatgct
     4801 taatcagtga ggcacctatc tcagcgatct gtctatttcg ttcatccata gttgcctgac
     4861 tccccgtcgt gtagataact acgatacggg agggcttacc atctggcccc agtgctgcaa
     4921 tgataccgcg agacccacgc tcaccggctc cagatttatc agcaataaac cagccagccg
     4981 gaagggccga gcgcagaagt ggtcctgcaa ctttatccgc ctccatccag tctattaatt
     5041 gttgccggga agctagagta agtagttcgc cagttaatag tttgcgcaac gttgttgcca
     5101 ttgctacagg catcgtggtg tcacgctcgt cgtttggtat ggcttcattc agctccggtt
     5161 cccaacgatc aaggcgagtt acatgatccc ccatgttgtg caaaaaagcg gttagctcct
     5221 tcggtcctcc gatcgttgtc agaagtaagt tggccgcagt gttatcactc atggttatgg
     5281 cagcactgca taattctctt actgtcatgc catccgtaag atgcttttct gtgactggtg
     5341 agtactcaac caagtcattc tgagaatagt gtatgcggcg accgagttgc tcttgcccgg
     5401 cgtcaatacg ggataatacc gcgccacata gcagaacttt aaaagtgctc atcattggaa
     5461 aacgttcttc ggggcgaaaa ctctcaagga tcttaccgct gttgagatcc agttcgatgt
     5521 aacccactcg tgcacccaac tgatcttcag catcttttac tttcaccagc gtttctgggt
     5581 gagcaaaaac aggaaggcaa aatgccgcaa aaaagggaat aagggcgaca cggaaatgtt
     5641 gaatactcat actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca
     5701 tgagcggata catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat
     5761 ttccccgaaa agtgccacct gacgtcgacg gatcgggaga tctcccgatc ccctatggtg
     5821 cactctcagt acaatctgct ctgatgccgc atagttaagc cagtatctgc tccctgcttg
     5881 tgtgttggag gtcgctgagt agtgcgcgag caaaatttaa gctacaacaa ggcaaggctt
     5941 gaccgacaat tgcatgaaga atctgcttag ggttaggcgt tttgcgctgc ttcgcgatgt
     6001 acgggccaga tatacgcgtt
//
