LOCUS       sequence-444343-        6596 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-2_Gav transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6596)
  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 6596)
  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-2_Gav
FEATURES             Location/Qualifiers
     source          1..6596
                     /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(2784..2802)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2785..2802)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2822..2839)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2828..3052
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      3098..3526
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3185..3204)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3395..3416
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3535..3555)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3540..3869
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3720..3855
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3782..3801
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    4035..4156
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(4072..4091)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     4126..4145
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4205..4221)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4205..4221)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4218..4240)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4229..4245
                     /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(4253..4259,4260..4277,4278..4283))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4298..4319
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4436..4453)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4607..5195)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4687..4706)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5366..6157,6158..6226))
                     /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     5989..6008
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6227..6331)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6406..6425)
                     /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 gagcaagtgc aaatgcgaag atggcgaatg tagcggcccg gccaaaaaaa agagctgcag
      781 ccgcttcaaa gaagattggc tgaaggaaat gtttgaaacc gatatgcctc atgccagcgg
      841 tcgtcaacag cgtaaaagct ggcgctattt ccctgtgagc tgcgaacgtg gtggcctgtg
      901 taccatatgc gttcgcgcag gtgcaaacag cgaatggacc acccgcaaaa aatggggcga
      961 ctggaaactg gactatctga aatgccatct gacccacaaa attcataccg acagcgtgat
     1021 gaaactgtgc aaccagagcc gcggcattct gaaaagcctg tttagcgaaa cgcgcgaaac
     1081 tcgcgaaaac cgcattgaac tgagcgaacg caaacgcagc aacgcggatg aagtgaaaat
     1141 cctgattgat aacgtgctgc tggcggtgaa actgaacctg agcatgctga gcgtgcagga
     1201 aatccacagc catgtgggca aatacgtgaa aatccctgaa agctggcgca gcaaaaacta
     1261 tgcgttcgag ttcatcgatt gcatcaacag cgtgattcat caggaagtgc tgaacgaaat
     1321 gcgcaccagc ccgttccata ccctgattgt ggacgaaagc accgatatta gcgtgagcaa
     1381 gatgctgatt ctgtacgtga aatttcgcac catggatgac tttacccacc gcactcgttt
     1441 tgccggcatt ctgaaactga ccgcgtgtga tagcagcgcg attgttgcgg cgattcgcca
     1501 gttctatgtg gatagcaacc tggacatgca gaaaatggtg atgttcacca gcgatggcgc
     1561 gagcgtgatg ttaggcaaac gcaatggcgt tgcggcagtt ttacgtcaga gcattccgca
     1621 tctgctggaa caacattgcg tggcgcatcg tgaggatctg ggcattgatg atgcgtggaa
     1681 agatgtgcct ctgatgcgct gcattgaaac cctgctgcgc accatctata ccctgtttag
     1741 ccgcagcagc gtgaaaaaga aaagcaagtt cgaggagctg agcaaagccg cggaatgcga
     1801 aactgttagc ttccgccctc tgaacgaagt tcgttggctg agccgccatt ttgcggttag
     1861 cgcgctgatg aagaactata ccgtgctgat ggattattgc gcgcagcagg tggcagagga
     1921 taacgatcct atccataagt actgcctgga taaactgcag aagcctgaat atcatgtggc
     1981 cctggcgatc ctgaatgatg tgctgggcga actggcagaa ctgtgtcgta tgctgcagcg
     2041 tagcagcctg actactgttg aggccttcca gtttgcgaaa gcgcgcatcg caaaactgcg
     2101 cagccagtat ttaggcgaaa ccgtgcattt tagcgaggat gtgcagactc tgctgagcag
     2161 ctttgaaagc ccggtggata ccaccagcgt gattcagttt attgaacaca cctgcgatca
     2221 tatggataac cgctttcctg aaggcgagct gaaagaatgg agcatctttg acaaggatag
     2281 ctttagcagc agcagcaacc gcaacgactt tagctttggc catgaagaac tgaagaccct
     2341 ggcgacccgt tatagcaccc tgctgaacaa aagcgtggaa accttcaaga aggatctgtg
     2401 caagcagtac gtggatttta aactgaccgt gagcgaaaag atgaagaccg gcgcgattaa
     2461 aacctttgaa gacatggtga cctacattct gcagcaggaa cagtttagcg aactgagcct
     2521 gctggtggat atttgcgcga ccttccaagc cagcagcgca gattgtgaac gcggctttag
     2581 cctgatgaac gccatcaaga ccaaaagccg caaccgcctg gaagtggata acctggataa
     2641 actgatgcgc atcaaactgc atctggccag cggcaagact gttaacctgg acaccgtgta
     2701 tagcgtgtgg aaaaccgaaa agggccgccg tgaaagcctg tggtaatgag cggccgctcg
     2761 agcatgcatc tagagggccc tattctatag tgtcacctaa atgctagagc tcgctgatca
     2821 gcctcgactg tgccttctag ttgccagcca tctgttgttt gcccctcccc cgtgccttcc
     2881 ttgaccctgg aaggtgccac tcccactgtc ctttcctaat aaaatgagga aattgcatcg
     2941 cattgtctga gtaggtgtca ttctattctg gggggtgggg tggggcagga cagcaagggg
     3001 gaggattggg aagacaatag caggcatgct ggggatgcgg tgggctctat ggcttctgag
     3061 gcggaaagaa ccagctgggg ctctaggggg tatccccacg cgccctgtag cggcgcatta
     3121 agcgcggcgg gtgtggtggt tacgcgcagc gtgaccgcta cacttgccag cgccctagcg
     3181 cccgctcctt tcgctttctt cccttccttt ctcgccacgt tcgccggctt tccccgtcaa
     3241 gctctaaatc gggggctccc tttagggttc cgatttagtg ctttacggca cctcgacccc
     3301 aaaaaacttg attagggtga tggttcacgt agtgggccat cgccctgata gacggttttt
     3361 cgccctttga cgttggagtc cacgttcttt aatagtggac tcttgttcca aactggaaca
     3421 acactcaacc ctatctcggt ctattctttt gatttataag ggattttgcc gatttcggcc
     3481 tattggttaa aaaatgagct gatttaacaa aaatttaacg cgaattaatt ctgtggaatg
     3541 tgtgtcagtt agggtgtgga aagtccccag gctccccagc aggcagaagt atgcaaagca
     3601 tgcatctcaa ttagtcagca accaggtgtg gaaagtcccc aggctcccca gcaggcagaa
     3661 gtatgcaaag catgcatctc aattagtcag caaccatagt cccgccccta actccgccca
     3721 tcccgcccct aactccgccc agttccgccc attctccgcc ccatggctga ctaatttttt
     3781 ttatttatgc agaggccgag gccgcctctg cctctgagct attccagaag tagtgaggag
     3841 gcttttttgg aggcctaggc ttttgcaaaa agctccccga aatgaccgac caagcgacgc
     3901 ccaacctgcc atcacgagat ttcgattcca ccgccgcctt ctatgaaagg ttgggcttcg
     3961 gaatcgtttt ccgggacgcc ggctggatga tcctccagcg cggggatctc atgctggagt
     4021 tcttcgccca ccccaacttg tttattgcag cttataatgg ttacaaataa agcaatagca
     4081 tcacaaattt cacaaataaa gcattttttt cactgcattc tagttgtggt ttgtccaaac
     4141 tcatcaatgt atcttatcat gtctgtatac cgtcgacctc tagctagagc ttggcgtaat
     4201 catggtcata gctgtttcct gtgtgaaatt gttatccgct cacaattcca cacaacatac
     4261 gagccggaag cataaagtgt aaagcctggg gtgcctaatg agtgagctaa ctcacattaa
     4321 ttgcgttgcg ctcactgccc gctttccagt cgggaaacct gtcgtgccag ctgcattaat
     4381 gaatcggcca acgcgcgggg agaggcggtt tgcgtattgg gcgctcttcc gcttcctcgc
     4441 tcactgactc gctgcgctcg gtcgttcggc tgcggcgagc ggtatcagct cactcaaagg
     4501 cggtaatacg gttatccaca gaatcagggg ataacgcagg aaagaacatg tgagcaaaag
     4561 gccagcaaaa ggccaggaac cgtaaaaagg ccgcgttgct ggcgtttttc cataggctcc
     4621 gcccccctga cgagcatcac aaaaatcgac gctcaagtca gaggtggcga aacccgacag
     4681 gactataaag ataccaggcg tttccccctg gaagctccct cgtgcgctct cctgttccga
     4741 ccctgccgct taccggatac ctgtccgcct ttctcccttc gggaagcgtg gcgctttctc
     4801 atagctcacg ctgtaggtat ctcagttcgg tgtaggtcgt tcgctccaag ctgggctgtg
     4861 tgcacgaacc ccccgttcag cccgaccgct gcgccttatc cggtaactat cgtcttgagt
     4921 ccaacccggt aagacacgac ttatcgccac tggcagcagc cactggtaac aggattagca
     4981 gagcgaggta tgtaggcggt gctacagagt tcttgaagtg gtggcctaac tacggctaca
     5041 ctagaagaac agtatttggt atctgcgctc tgctgaagcc agttaccttc ggaaaaagag
     5101 ttggtagctc ttgatccggc aaacaaacca ccgctggtag cggtggtttt tttgtttgca
     5161 agcagcagat tacgcgcaga aaaaaaggat ctcaagaaga tcctttgatc ttttctacgg
     5221 ggtctgacgc tcagtggaac gaaaactcac gttaagggat tttggtcatg agattatcaa
     5281 aaaggatctt cacctagatc cttttaaatt aaaaatgaag ttttaaatca atctaaagta
     5341 tatatgagta aacttggtct gacagttacc aatgcttaat cagtgaggca cctatctcag
     5401 cgatctgtct atttcgttca tccatagttg cctgactccc cgtcgtgtag ataactacga
     5461 tacgggaggg cttaccatct ggccccagtg ctgcaatgat accgcgagac ccacgctcac
     5521 cggctccaga tttatcagca ataaaccagc cagccggaag ggccgagcgc agaagtggtc
     5581 ctgcaacttt atccgcctcc atccagtcta ttaattgttg ccgggaagct agagtaagta
     5641 gttcgccagt taatagtttg cgcaacgttg ttgccattgc tacaggcatc gtggtgtcac
     5701 gctcgtcgtt tggtatggct tcattcagct ccggttccca acgatcaagg cgagttacat
     5761 gatcccccat gttgtgcaaa aaagcggtta gctccttcgg tcctccgatc gttgtcagaa
     5821 gtaagttggc cgcagtgtta tcactcatgg ttatggcagc actgcataat tctcttactg
     5881 tcatgccatc cgtaagatgc ttttctgtga ctggtgagta ctcaaccaag tcattctgag
     5941 aatagtgtat gcggcgaccg agttgctctt gcccggcgtc aatacgggat aataccgcgc
     6001 cacatagcag aactttaaaa gtgctcatca ttggaaaacg ttcttcgggg cgaaaactct
     6061 caaggatctt accgctgttg agatccagtt cgatgtaacc cactcgtgca cccaactgat
     6121 cttcagcatc ttttactttc accagcgttt ctgggtgagc aaaaacagga aggcaaaatg
     6181 ccgcaaaaaa gggaataagg gcgacacgga aatgttgaat actcatactc ttcctttttc
     6241 aatattattg aagcatttat cagggttatt gtctcatgag cggatacata tttgaatgta
     6301 tttagaaaaa taaacaaata ggggttccgc gcacatttcc ccgaaaagtg ccacctgacg
     6361 tcgacggatc gggagatctc ccgatcccct atggtgcact ctcagtacaa tctgctctga
     6421 tgccgcatag ttaagccagt atctgctccc tgcttgtgtg ttggaggtcg ctgagtagtg
     6481 cgcgagcaaa atttaagcta caacaaggca aggcttgacc gacaattgca tgaagaatct
     6541 gcttagggtt aggcgttttg cgctgcttcg cgatgtacgg gccagatata cgcgtt
//
