LOCUS       sequence-444287-        6419 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  Mammalian expression vector for expression of hAT-3_AMi transposase 
            (CMV promoter).
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6419)
  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 6419)
  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_AMi
FEATURES             Location/Qualifiers
     source          1..6419
                     /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(2607..2625)
                     /label=SP6 promoter
                     /note="promoter for bacteriophage SP6 RNA polymerase"
     primer_bind     complement(2608..2625)
                     /label=SP6
                     /note="SP6 promoter, forward primer"
     primer_bind     complement(2645..2662)
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
     polyA_signal    2651..2875
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     rep_origin      2921..3349
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(3008..3027)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3218..3239
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     complement(3358..3378)
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     promoter        3363..3692
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      3543..3678
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     3605..3624
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     polyA_signal    3858..3979
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3895..3914)
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     primer_bind     3949..3968
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     complement(4028..4044)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     complement(4028..4044)
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     primer_bind     complement(4041..4063)
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     protein_bind    4052..4068
                     /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(4076..4082,4083..4100,4101..4106))
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4121..4142
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     primer_bind     complement(4259..4276)
                     /label=L4440
                     /note="L4440 vector, forward primer"
     rep_origin      complement(4430..5018)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     complement(4510..4529)
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     CDS             complement(join(5189..5980,5981..6049))
                     /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     5812..5831
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     promoter        complement(6050..6154)
                     /gene="bla"
                     /label=AmpR promoter
     primer_bind     complement(6229..6248)
                     /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 gtgcaagttt gtgcagcgct gcaaattatc gggccgcatt gattcagttc tgagcgcgca
      781 gtttgaaaaa gaatgcagct actggcgcaa ggttctgcgt cgtgtggtgg cgaccgttaa
      841 actgctgagc agcttaggtc tgcctttacg cggccacgat gaaagctcac tgagcaaccg
      901 caaaggcaac ttcctgacct gcctggaata cctgagcgag tttgatgact tcctgaaaga
      961 gcatctgaag aactaccagt attgcggcag cggcaagacc aactttctga cccatcagac
     1021 ctacgatgag tttatcacca tcatggcgga gcgtctgcgt aaccagttca ccgatgaagt
     1081 gaaagacgcg aaatactaca gcattattgt ggacagcacc cctgatgtga gccatgtgga
     1141 tcagctgacc ctgattctgc gttacgtgac cggtaacggc gaagtggtgg aacgctttct
     1201 gtgctttgtg cctctgaaga gccataccag cgagtgtctg gaaaccaccg tgctggagat
     1261 tattgcgaac ctgggcctgg acattaagaa ctgccatggc cagagctttg acaacgccgc
     1321 gaacatggcg ggtaaatata gcggtctgca ggcgcgttta aacaatgcaa gcccgagcgc
     1381 gttatttatt ccttgcagca gccatagcct gaacctgatt tgtaacgcgg cggcggaatc
     1441 atgtgaaggc gcaacccact tcttctattt tgtgcagaac gtgtatgcgt tttttagcgt
     1501 gagcacccat cgctggagca tgctgcaaag ccatctggag caggccaacg aaaaacacct
     1561 gaccgtgaag cgcatatgcg atacccgttg gtcagcgcgt gcggatgcgg ttttagcgct
     1621 gaaaatgagc tacgacgaca tcaaaaaaac cctgctggat atcgcgacca gcaactcaga
     1681 aaagccgcgc gcgaaaaccg aagcgaaaag cctggccgag aagtttgaga aatatgacat
     1741 cgccgtgttt accctgttat ggaaccgcct gctgcaacgc attaacgcgg tgagcaaact
     1801 gctgcagaaa gtggatacca acctgctgaa tgcggcgcaa ttactggcga gcgtgaaaag
     1861 cttcgtgatg gaggtgcgca acgattatag cagcgtggaa gccgaagcgt taaccctgac
     1921 cgaaaacatt ggcagcagcg acattagcga cgaaaaacgc aagaaaagcc gcaaactgtt
     1981 tttcgatgag actcgcgata acgaaattaa cctgaagggc aaagagaaga ttatcgtgga
     2041 aaccgtgaac gtgatttgcg ataccattat tgtgcagctg cagagccgtg gcaagagcct
     2101 gaaaaaaacc gtgaacctgt tttgcgtgtt tttcgatcgc agcatggatg aaaacgcgaa
     2161 aagccactac agcaaaaaac tgagcgagtt ttaccgcgaa gatattgata ccaacctgtt
     2221 tgaggatgag gtgaagcact tcatccactt cattgaaaac gacgaggtgt gtgcgtcacg
     2281 tagccctgtg gatttatatc gcctggtgcg cgatggtctg caagccacct ttccgaacgt
     2341 ggaaaccatc ctgaaaatct ttctgaccat tcctgtgacc aacgcatcag gcgaacgcag
     2401 cttttcagtg ctgaagcgcg tgaagaacta cctgcgcaac accatgagcc aggaacatct
     2461 gaccagcagc gcgattctga ccattgaaag cgccagcctg cagaacatta cctatgacga
     2521 tctgatcgac gatttcgtga agaaaaagtg ccgcaagaag gcgatctaat gagcggccgc
     2581 tcgagcatgc atctagaggg ccctattcta tagtgtcacc taaatgctag agctcgctga
     2641 tcagcctcga ctgtgccttc tagttgccag ccatctgttg tttgcccctc ccccgtgcct
     2701 tccttgaccc tggaaggtgc cactcccact gtcctttcct aataaaatga ggaaattgca
     2761 tcgcattgtc tgagtaggtg tcattctatt ctggggggtg gggtggggca ggacagcaag
     2821 ggggaggatt gggaagacaa tagcaggcat gctggggatg cggtgggctc tatggcttct
     2881 gaggcggaaa gaaccagctg gggctctagg gggtatcccc acgcgccctg tagcggcgca
     2941 ttaagcgcgg cgggtgtggt ggttacgcgc agcgtgaccg ctacacttgc cagcgcccta
     3001 gcgcccgctc ctttcgcttt cttcccttcc tttctcgcca cgttcgccgg ctttccccgt
     3061 caagctctaa atcgggggct ccctttaggg ttccgattta gtgctttacg gcacctcgac
     3121 cccaaaaaac ttgattaggg tgatggttca cgtagtgggc catcgccctg atagacggtt
     3181 tttcgccctt tgacgttgga gtccacgttc tttaatagtg gactcttgtt ccaaactgga
     3241 acaacactca accctatctc ggtctattct tttgatttat aagggatttt gccgatttcg
     3301 gcctattggt taaaaaatga gctgatttaa caaaaattta acgcgaatta attctgtgga
     3361 atgtgtgtca gttagggtgt ggaaagtccc caggctcccc agcaggcaga agtatgcaaa
     3421 gcatgcatct caattagtca gcaaccaggt gtggaaagtc cccaggctcc ccagcaggca
     3481 gaagtatgca aagcatgcat ctcaattagt cagcaaccat agtcccgccc ctaactccgc
     3541 ccatcccgcc cctaactccg cccagttccg cccattctcc gccccatggc tgactaattt
     3601 tttttattta tgcagaggcc gaggccgcct ctgcctctga gctattccag aagtagtgag
     3661 gaggcttttt tggaggccta ggcttttgca aaaagctccc cgaaatgacc gaccaagcga
     3721 cgcccaacct gccatcacga gatttcgatt ccaccgccgc cttctatgaa aggttgggct
     3781 tcggaatcgt tttccgggac gccggctgga tgatcctcca gcgcggggat ctcatgctgg
     3841 agttcttcgc ccaccccaac ttgtttattg cagcttataa tggttacaaa taaagcaata
     3901 gcatcacaaa tttcacaaat aaagcatttt tttcactgca ttctagttgt ggtttgtcca
     3961 aactcatcaa tgtatcttat catgtctgta taccgtcgac ctctagctag agcttggcgt
     4021 aatcatggtc atagctgttt cctgtgtgaa attgttatcc gctcacaatt ccacacaaca
     4081 tacgagccgg aagcataaag tgtaaagcct ggggtgccta atgagtgagc taactcacat
     4141 taattgcgtt gcgctcactg cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt
     4201 aatgaatcgg ccaacgcgcg gggagaggcg gtttgcgtat tgggcgctct tccgcttcct
     4261 cgctcactga ctcgctgcgc tcggtcgttc ggctgcggcg agcggtatca gctcactcaa
     4321 aggcggtaat acggttatcc acagaatcag gggataacgc aggaaagaac atgtgagcaa
     4381 aaggccagca aaaggccagg aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc
     4441 tccgcccccc tgacgagcat cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga
     4501 caggactata aagataccag gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc
     4561 cgaccctgcc gcttaccgga tacctgtccg cctttctccc ttcgggaagc gtggcgcttt
     4621 ctcatagctc acgctgtagg tatctcagtt cggtgtaggt cgttcgctcc aagctgggct
     4681 gtgtgcacga accccccgtt cagcccgacc gctgcgcctt atccggtaac tatcgtcttg
     4741 agtccaaccc ggtaagacac gacttatcgc cactggcagc agccactggt aacaggatta
     4801 gcagagcgag gtatgtaggc ggtgctacag agttcttgaa gtggtggcct aactacggct
     4861 acactagaag aacagtattt ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa
     4921 gagttggtag ctcttgatcc ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt
     4981 gcaagcagca gattacgcgc agaaaaaaag gatctcaaga agatcctttg atcttttcta
     5041 cggggtctga cgctcagtgg aacgaaaact cacgttaagg gattttggtc atgagattat
     5101 caaaaaggat cttcacctag atccttttaa attaaaaatg aagttttaaa tcaatctaaa
     5161 gtatatatga gtaaacttgg tctgacagtt accaatgctt aatcagtgag gcacctatct
     5221 cagcgatctg tctatttcgt tcatccatag ttgcctgact ccccgtcgtg tagataacta
     5281 cgatacggga gggcttacca tctggcccca gtgctgcaat gataccgcga gacccacgct
     5341 caccggctcc agatttatca gcaataaacc agccagccgg aagggccgag cgcagaagtg
     5401 gtcctgcaac tttatccgcc tccatccagt ctattaattg ttgccgggaa gctagagtaa
     5461 gtagttcgcc agttaatagt ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt
     5521 cacgctcgtc gtttggtatg gcttcattca gctccggttc ccaacgatca aggcgagtta
     5581 catgatcccc catgttgtgc aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca
     5641 gaagtaagtt ggccgcagtg ttatcactca tggttatggc agcactgcat aattctctta
     5701 ctgtcatgcc atccgtaaga tgcttttctg tgactggtga gtactcaacc aagtcattct
     5761 gagaatagtg tatgcggcga ccgagttgct cttgcccggc gtcaatacgg gataataccg
     5821 cgccacatag cagaacttta aaagtgctca tcattggaaa acgttcttcg gggcgaaaac
     5881 tctcaaggat cttaccgctg ttgagatcca gttcgatgta acccactcgt gcacccaact
     5941 gatcttcagc atcttttact ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa
     6001 atgccgcaaa aaagggaata agggcgacac ggaaatgttg aatactcata ctcttccttt
     6061 ttcaatatta ttgaagcatt tatcagggtt attgtctcat gagcggatac atatttgaat
     6121 gtatttagaa aaataaacaa ataggggttc cgcgcacatt tccccgaaaa gtgccacctg
     6181 acgtcgacgg atcgggagat ctcccgatcc cctatggtgc actctcagta caatctgctc
     6241 tgatgccgca tagttaagcc agtatctgct ccctgcttgt gtgttggagg tcgctgagta
     6301 gtgcgcgagc aaaatttaag ctacaacaag gcaaggcttg accgacaatt gcatgaagaa
     6361 tctgcttagg gttaggcgtt ttgcgctgct tcgcgatgta cgggccagat atacgcgtt
//
