LOCUS       sequence-13972-e        5357 bp    DNA     circular SYN 15-DEC-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 5357)
  AUTHORS   Moss B, Elroy-Stein O, Mizukami T, Alexander WA, Fuerst TR
  TITLE     Product review. New mammalian expression vectors.
  JOURNAL   Nature. 1990 Nov 1. 348(6296):91-2.
  PUBMED    2234068
REFERENCE   2  (bases 1 to 5357)
  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"; journalName: "Nature. 
            1990 Nov 1. 348(6296):91-2."
COMMENT     Sequence Label: pTM1
FEATURES             Location/Qualifiers
     source          1..5357
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     primer_bind     794..813
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        794..812
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     misc_feature    851..1422
                     /label=IRES
                     /note="internal ribosome entry site (IRES) of the 
                     encephalomyocarditis virus (EMCV)"
     primer_bind     complement(1018..1035)
                     /label=IRES reverse
                     /note="IRES internal ribosome entry site, reverse primer. 
                     Also called pCDH-rev"
     primer_bind     1245..1264
                     /label=IRES-F
                     /note="IRES internal ribosome entry site, forward primer"
     primer_bind     complement(1542..1560)
                     /label=T7 Term
                     /note="T7 terminator, reverse primer"
     terminator      1556..1603
                     /label=T7 terminator
                     /note="transcription terminator for bacteriophage T7 RNA 
                     polymerase"
     primer_bind     complement(2583..2605)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      2735..3190
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(2822..2841)
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     primer_bind     3032..3053
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     promoter        3219..3323
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(3324..3392,3393..4184)
                     /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     complement(3542..3561)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      4355..4943
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     4844..4863
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     5097..5114
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    5231..5252
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(5267..5272,5273..5290,5291..5297)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    5305..5321
                     /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)."
     primer_bind     5310..5332
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     5329..5345
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     5329..5345
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
ORIGIN
        1 aagcttttgc gatcaataaa tggatcacaa ccagtatctc ttaacgatgt tcttcgcaga
       61 tgatgattca ttttttaagt atttggctag tcaagatgat gaatcttcat tatctgatat
      121 attgcaaatc actcaatatc tagactttct gttattatta ttgatccaat caaaaaataa
      181 attagaagcc gtgggtcatt gttatgaatc tctttcagag gaatacagac aattgacaaa
      241 attcacagac tttcaagatt ttaaaaaact gtttaacaag gtccctattg ttacagatgg
      301 aagggtcaaa cttaataaag gatatttgtt cgactttgtg attagtttga tgcgattcaa
      361 aaaagaatcc tctctagcta ccaccgcaat agatcctgtt agatacatag atcctcgtcg
      421 caatatcgca ttttctaacg tgatggatat attaaagtcg aataaagtga acaataatta
      481 attctttatt gtcatcatga acggcggaca tattcagttg ataatcggcc ccatgttttc
      541 aggtaaaagt acagaattaa ttagacgagt tagacgttat caaatagctc aatataaatg
      601 cgtgactata aaatattcta acgataatag atacggaacg ggactatgga cgcatgataa
      661 gaataatttt gaagcattgg aagcaactaa actatgtgat gtcttggaat caattacaga
      721 tttctccgtg ataggtatcg atgaaggaca gttctttcca gacattgttg aattgatctc
      781 gatcccgcga aattaatacg actcactata gggagaccac aacggtttcc ctctagcggg
      841 atcaattccg cccctctccc tccccccccc ctaacgttac tggccgaagc cgcttggaat
      901 aaggccggtg tgcgtttgtc tatatgttat tttccaccat attgccgtct tttggcaatg
      961 tgagggcccg gaaacctggc cctgtcttct tgacgagcat tcctaggggt ctttcccctc
     1021 tcgccaaagg aatgcaaggt ctgttgaatg tcgtgaagga agcagttcct ctggaagctt
     1081 cttgaagaca aacaacgtct gtagcgaccc tttgcaggca gcggaacccc ccacctggcg
     1141 acaggtgcct ctgcggccaa aagccacgtg tataagatac acctgcaaag gcggcacaac
     1201 cccagtgcca cgttgtgagt tggatagttg tggaaagagt caaatggctc tcctcaagcg
     1261 tattcaacaa ggggctgaag gatgcccaga aggtacccca ttgtatggga tctgatctgg
     1321 ggcctcggtg cacatgcttt acatgtgttt agtcgaggtt aaaaaacgtc taggcccccc
     1381 gaaccacggg gacgtggttt tcctttgaaa aacacgataa taccatggga attccccggg
     1441 gagctcacta gtggatccct gcagctcgag aggcctaatt aattaagtcg acgatccggc
     1501 tgctaacaaa gcccgaaagg aagctgagtt ggctgctgcc accgctgagc aataactagc
     1561 ataacccctt ggggcctcta aacgggtctt gaggggtttt ttgctgaaag gaggaactat
     1621 atccggatcg agatcaattc tgtgagcgta tggcaaacga aggaaaaata gttatagtag
     1681 ccgcactcga tgggacattt caacgtaaac cgtttaataa tattttgaat cttattccat
     1741 tatctgaaat ggtggtaaaa ctaactgctg tgtgtatgaa atgctttaag gaggcttcct
     1801 tttctaaacg attgggtgag gaaaccgaga tagaaataat aggaggtaat gatatgtatc
     1861 aatcggtgtg tagaaagtgt tacatcgact cataatatta tattttttat ctaaaaaact
     1921 aaaaataaac attgattaaa ttttaatata atacttaaaa atggatgttg tgtcgttaga
     1981 taaaccgttt atgtattttg aggaaattga taatgagtta gattacgaac cagaaagtgc
     2041 aaatgaggtc gcaaaaaaac tgccgtatca aggacagtta aaactattac taggagaatt
     2101 attttttctt agtaagttac agcgacacgg tatattagat ggtgccaccg tagtgtatat
     2161 aggatctgct cccggtacac atatacgtta tttgagagat catttctata atttaggagt
     2221 gatcatcaaa tggatgctaa ttgacggccg ccatcatgat cctattttaa atggattgcg
     2281 tgatgtgact ctagtgactc ggttcgttga tgaggaatat ctacgatcca tcaaaaaaca
     2341 actgcatcct tctaagatta ttttaatttc tgatgtgaga tccaaacgag gaggaaatga
     2401 acctagtacg gcggatttac taagtaatta cgctctacaa aatgtcatga ttagtatttt
     2461 aaaccccgtg gcgtctagtc ttaaatggag atgcccgttt ccagatcaat ggatcaagga
     2521 cttttatatc ccacacggta ataaaatgtt acaacctttt gctccttcat attcagggcc
     2581 gtcgttttac aacgtcgtga ctgggaaaac cctggcgtta cccaacttaa tcgccttgca
     2641 gcacatcccc ctttcgccag ctggcgtaat agcgaagagg cccgcaccga tcgcccttcc
     2701 caacagttgc gcagcctgaa tggcgaatgg cgcgacgcgc cctgtagcgg cgcattaagc
     2761 gcggcgggtg tggtggttac gcgcagcgtg accgctacac ttgccagcgc cctagcgccc
     2821 gctcctttcg ctttcttccc ttcctttctc gccacgttcg ccggctttcc ccgtcaagct
     2881 ctaaatcggg ggctcccttt agggttccga tttagtgctt tacggcacct cgaccccaaa
     2941 aaacttgatt agggtgatgg ttcacgtagt gggccatcgc cctgatagac ggtttttcgc
     3001 cctttgacgt tggagtccac gttctttaat agtggactct tgttccaaac tggaacaaca
     3061 ctcaacccta tctcggtcta ttcttttgat ttataaggga ttttgccgat ttcggcctat
     3121 tggttaaaaa atgagctgat ttaacaaaaa tttaacgcga attttaacaa aatattaacg
     3181 tttacaattt cccaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat
     3241 ttttctaaat acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc
     3301 aataatattg aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct
     3361 tttttgcggc attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag
     3421 atgctgaaga tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta
     3481 agatccttga gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc
     3541 tgctatgtgg cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca
     3601 tacactattc tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg
     3661 atggcatgac agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg
     3721 ccaacttact tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca
     3781 tgggggatca tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa
     3841 acgacgagcg tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa
     3901 ctggcgaact acttactcta gcttcccggc aacaattaat agactggatg gaggcggata
     3961 aagttgcagg accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat
     4021 ctggagccgg tgagcgtggg tctcgcggta tcattgcagc actggggcca gatggtaagc
     4081 cctcccgtat cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata
     4141 gacagatcgc tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt
     4201 actcatatat actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga
     4261 agatcctttt tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag
     4321 cgtcagaccc cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa
     4381 tctgctgctt gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag
     4441 agctaccaac tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg
     4501 tccttctagt gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat
     4561 acctcgctct gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta
     4621 ccgggttgga ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg
     4681 gttcgtgcac acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc
     4741 gtgagctatg agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa
     4801 gcggcagggt cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc
     4861 tttatagtcc tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt
     4921 caggggggcg gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct
     4981 tttgctggcc ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc
     5041 gtattaccgc ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg
     5101 agtcagtgag cgaggaagcg gaagagcgcc caatacgcaa accgcctctc cccgcgcgtt
     5161 ggccgattca ttaatgcagc tggcacgaca ggtttcccga ctggaaagcg ggcagtgagc
     5221 gcaacgcaat taatgtgagt tagctcactc attaggcacc ccaggcttta cactttatgc
     5281 ttccggctcg tatgttgtgt ggaattgtga gcggataaca atttcacaca ggaaacagct
     5341 atgaccatga ttacgcc
//
