LOCUS       sequence-450097-        7056 bp    DNA     circular SYN 15-APR-2025
DEFINITION  pcDNA-dTdT.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7056)
  AUTHORS   Carlson CK, Loveless TB, Milisavljevic M, Kelly PI, Mills JH, Tyo 
            KEJ, Liu CC
  TITLE     A massively parallel in vivo assay of TdT mutants yields variants 
            with altered nucleotide insertion biases.
  JOURNAL   bioRxiv [Preprint]. 2024 Jun 11:2024.06.11.598561. doi: 
            10.1101/2024.06.11.598561.
  PUBMED    38915690
REFERENCE   2  (bases 1 to 7056)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Apr 15, 2025 from SnapGene Server 8.0.1
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"; journalName: "bioRxiv 
            [Preprint]. 2024 Jun 11:2024.06.11.598561. doi: 
            10.1101/2024.06.11.598561."
COMMENT     Sequence Label: pTBL435
FEATURES             Location/Qualifiers
     source          1..7056
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     regulatory      666..675
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
                     /regulatory_class="other"
     primer_bind     complement(704..723)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(705..723)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     promoter        complement(768..971)
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early 
                     promoter"
     primer_bind     complement(797..817)
                     /label=CMV-F
                     /note="Human CMV immediate early promoter, forward primer"
     enhancer        972..1351
                     /label=CMV enhancer
                     /note="human cytomegalovirus immediate early enhancer"
     primer_bind     1523..1542
                     /label=pRS-marker
                     /note="pRS vectors, use to sequence yeast selectable 
                     marker"
     promoter        1617..1721
                     /gene="bla"
                     /label=AmpR promoter
     CDS             join(1722..1790,1791..2582)
                     /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(1940..1959)
                     /label=Amp-R
                     /note="Ampicillin resistance gene, reverse primer"
     rep_origin      2753..3341
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     3242..3261
                     /label=pBR322ori-F
                     /note="pBR322 origin, forward primer"
     primer_bind     3495..3512
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    3629..3650
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        join(3665..3670,3671..3688,3689..3695)
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    3703..3719
                     /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     3708..3730
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     3727..3743
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     3727..3743
                     /label=M13 Reverse
                     /note="In lacZ gene. Also called M13-rev"
     polyA_signal    3792..3913
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     primer_bind     complement(3803..3822)
                     /label=EBV-rev
                     /note="SV40 polyA terminator, reverse primer"
     primer_bind     3857..3876
                     /label=SV40pA-R
                     /note="SV40 polyA, reverse primer"
     promoter        complement(5104..5433)
                     /label=SV40 promoter
                     /note="SV40 enhancer and early promoter"
     rep_origin      5118..5253
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(5172..5191)
                     /label=SV40pro-F
                     /note="SV40 promoter/origin, forward primer"
     primer_bind     5418..5438
                     /label=pBABE 3'
                     /note="SV40 enhancer, reverse primer for pBABE vectors"
     rep_origin      complement(5447..5875)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     primer_bind     complement(5557..5578)
                     /label=F1ori-F
                     /note="F1 origin, forward primer"
     primer_bind     5769..5788
                     /label=F1ori-R
                     /note="F1 origin, reverse primer"
     polyA_signal    5921..6145
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     primer_bind     6134..6151
                     /label=BGH-rev
                     /note="Bovine growth hormone terminator, reverse primer. 
                     Also called BGH reverse"
ORIGIN
        1 acccttcacc ttggacccca ggcagggaat tccttctgtg tccttcatac tgatgattgt
       61 gaatggcaga gatttcaata cagaagctgc tctcataaat gtcacacagg agtcttcatt
      121 ttctctaaac tcacagtttt cagccagtat atcaaaggca tccgtgaata tctggttaca
      181 gttgtttaaa gtggttcttc tctgacacgc atactgggag atcttttgta cagcaattgg
      241 tggagtcttc ggggggcctg ggttggtgct atctgaatag tctcttctca caacaagctg
      301 gtgttttcct gtcatttcca ccggtttccc tgctcctatg cattcgatca gccaggagac
      361 atcgaggagc tctggttgtg agctgacttg tactttctgt gcttgaagcc actccagaac
      421 atccgaaccc gagttgttct ctgctacaat gtgggtgaca gaatcactga gctcattttc
      481 aaccctgaac cctttcctgc gggccagctc catgaggaac gctctgcggg tggttcccat
      541 tttcttctcc aaaatgaaga cgaccaaatc ttgaaatttg atgtcttgag gagaggaggc
      601 catcaaggca cccgtctgcc ggggtctctt cttccgaggg ctcaagtggg acgctcgtgg
      661 tggatccatg gtggcgaccg gtagcgctag ccagcttggg tctccctata gtgagtcgta
      721 ttaatttcga taagccagta agcagtgggt tctctagtta gccagagagc tctgcttata
      781 tagacctccc accgtacacg cctaccgccc atttgcgtca atggggcgga gttgttacga
      841 cattttggaa agtcccgttg attttggtgc caaaacaaac tcccattgac gtcaatgggg
      901 tggagacttg gaaatccccg tgagtcaaac cgctatccac gcccattgat gtactgccaa
      961 aaccgcatca ccatggtaat agcgatgact aatacgtaga tgtactgcca agtaggaaag
     1021 tcccataagg tcatgtactg ggcataatgc caggcgggcc atttaccgtc attgacgtca
     1081 atagggggcg tacttggcat atgatacact tgatgtactg ccaagtgggc agtttaccgt
     1141 aaatactcca cccattgacg tcaatggaaa gtccctattg gcgttactat gggaacatac
     1201 gtcattattg acgtcaatgg gcgggggtcg ttgggcggtc agccaggcgg gccatttacc
     1261 gtaagttatg taacgcggaa ctccatatat gggctatgaa ctaatgaccc cgtaattgat
     1321 tactattaat aactagtcaa taatcaatgt caacgcgtat atctggcccg tacatcgcga
     1381 agcagcgcaa aacgcctaac cctaagcaga ttcttcatgc aattgtcggt caagccttgc
     1441 cttgttgtag cttaaatttt gctcgcgcac tactcagcga cctccaacac acaggcaggg
     1501 agcagatact ggcttaacta tgcggcatca gagcagattg tactgagagt gcaccatagg
     1561 ggatcgggag atctcccgat ccgtcgacgt caggtggcac ttttcgggga aatgtgcgcg
     1621 gaacccctat ttgtttattt ttctaaatac attcaaatat gtatccgctc atgagacaat
     1681 aaccctgata aatgcttcaa taatattgaa aaaggaagag tatgagtatt caacatttcc
     1741 gtgtcgccct tattcccttt tttgcggcat tttgccttcc tgtttttgct cacccagaaa
     1801 cgctggtgaa agtaaaagat gctgaagatc agttgggtgc acgagtgggt tacatcgaac
     1861 tggatctcaa cagcggtaag atccttgaga gttttcgccc cgaagaacgt tttccaatga
     1921 tgagcacttt taaagttctg ctatgtggcg cggtattatc ccgtattgac gccgggcaag
     1981 agcaactcgg tcgccgcata cactattctc agaatgactt ggttgagtac tcaccagtca
     2041 cagaaaagca tcttacggat ggcatgacag taagagaatt atgcagtgct gccataacca
     2101 tgagtgataa cactgcggcc aacttacttc tgacaacgat cggaggaccg aaggagctaa
     2161 ccgctttttt gcacaacatg ggggatcatg taactcgcct tgatcgttgg gaaccggagc
     2221 tgaatgaagc cataccaaac gacgagcgtg acaccacgat gcctgtagca atggcaacaa
     2281 cgttgcgcaa actattaact ggcgaactac ttactctagc ttcccggcaa caattaatag
     2341 actggatgga ggcggataaa gttgcaggac cacttctgcg ctcggccctt ccggctggct
     2401 ggtttattgc tgataaatct ggagccggtg agcgtgggtc tcgcggtatc attgcagcac
     2461 tggggccaga tggtaagccc tcccgtatcg tagttatcta cacgacgggg agtcaggcaa
     2521 ctatggatga acgaaataga cagatcgctg agataggtgc ctcactgatt aagcattggt
     2581 aactgtcaga ccaagtttac tcatatatac tttagattga tttaaaactt catttttaat
     2641 ttaaaaggat ctaggtgaag atcctttttg ataatctcat gaccaaaatc ccttaacgtg
     2701 agttttcgtt ccactgagcg tcagaccccg tagaaaagat caaaggatct tcttgagatc
     2761 ctttttttct gcgcgtaatc tgctgcttgc aaacaaaaaa accaccgcta ccagcggtgg
     2821 tttgtttgcc ggatcaagag ctaccaactc tttttccgaa ggtaactggc ttcagcagag
     2881 cgcagatacc aaatactgtt cttctagtgt agccgtagtt aggccaccac ttcaagaact
     2941 ctgtagcacc gcctacatac ctcgctctgc taatcctgtt accagtggct gctgccagtg
     3001 gcgataagtc gtgtcttacc gggttggact caagacgata gttaccggat aaggcgcagc
     3061 ggtcgggctg aacggggggt tcgtgcacac agcccagctt ggagcgaacg acctacaccg
     3121 aactgagata cctacagcgt gagctatgag aaagcgccac gcttcccgaa gggagaaagg
     3181 cggacaggta tccggtaagc ggcagggtcg gaacaggaga gcgcacgagg gagcttccag
     3241 ggggaaacgc ctggtatctt tatagtcctg tcgggtttcg ccacctctga cttgagcgtc
     3301 gatttttgtg atgctcgtca ggggggcgga gcctatggaa aaacgccagc aacgcggcct
     3361 ttttacggtt cctggccttt tgctggcctt ttgctcacat gttctttcct gcgttatccc
     3421 ctgattctgt ggataaccgt attaccgcct ttgagtgagc tgataccgct cgccgcagcc
     3481 gaacgaccga gcgcagcgag tcagtgagcg aggaagcgga agagcgccca atacgcaaac
     3541 cgcctctccc cgcgcgttgg ccgattcatt aatgcagctg gcacgacagg tttcccgact
     3601 ggaaagcggg cagtgagcgc aacgcaatta atgtgagtta gctcactcat taggcacccc
     3661 aggctttaca ctttatgctt ccggctcgta tgttgtgtgg aattgtgagc ggataacaat
     3721 ttcacacagg aaacagctat gaccatgatt acgccaagct ctagctagag gtcgacggta
     3781 tacagacatg ataagataca ttgatgagtt tggacaaacc acaactagaa tgcagtgaaa
     3841 aaaatgcttt atttgtgaaa tttgtgatgc tattgcttta tttgtaacca ttataagctg
     3901 caataaacaa gttggggtgg gcgaagaact ccagcatgag atccccgcgc tggaggatca
     3961 tccagccggc gtcccggaaa acgattccga agcccaacct ttcatagaag gcggcggtgg
     4021 aatcgaaatc tcgtgatggc aggttgggcg tcgcttggtc ggtcatttcg aaccccagag
     4081 tcccgcctat tcctttgccc tcggacgagt gctggggcgt cggtttccac tatcggcgag
     4141 tacttctaca cagccatcgg tccagacggc cgcgcttctg cgggcgattt gtgtacgccc
     4201 gacagtcccg gctccggatc ggacgattgc gtcgcatcga ccctgcgccc aagctgcatc
     4261 atcgaaattg ccgtcaacca agctctgata gagttggtca agaccaatgc ggagcatata
     4321 cgcccggagc cgcggcgatc ctgcaagctc cggatgcctc cgctcgaagt agcgcgtctg
     4381 ctgctccata caagccaacc acggcctcca gaagaagatg ttggcgacct cgtattggga
     4441 atccccgaac atcgcctcgc tccagtcaat gaccgctgtt atgcggccat tgtccgtcag
     4501 gacattgttg gagccgaaat ccgcgtgcac gaggtgccgg acttcggggc agtcctcggc
     4561 ccaaagcatc agctcatcga gagcctgcgc gacggacgca ctgacggtgt cgtccatcac
     4621 agtttgccag tgatacacat ggggatcagc aatcgcgcat atgaaatcac gccatgtagt
     4681 gtattgaccg attccttgcg gtccgaatgg gccgaacccg ctcgtctggc taagatcggc
     4741 cgcagcgatc gcatccatgg cctccgcgac cggttgcaga acagcgggca gttcggtttc
     4801 aggcaggtct tgcaacgtga caccctgtgc acggcgggag atgcaatagg tcaggctctc
     4861 gctaaattcc ccaatgtcaa gcacttccgg aatcgggagc gcggccgatg caaagtgccg
     4921 ataaacataa cgatctttgt agaaaccatc ggcgcagcta tttacccgca ggacatatcc
     4981 acgccctcct acatcgaagc tgaaagcacg agattcttcg ccctccgaga gctgcatgcg
     5041 aaacgatcct catcctgtct cttgatcaga tccgaaaatg gatatacaag ctcccgggag
     5101 ctttttgcaa aagcctaggc ctccaaaaaa gcctcctcac tacttctgga atagctcaga
     5161 ggcagaggcg gcctcggcct ctgcataaat aaaaaaaatt agtcagccat ggggcggaga
     5221 atgggcggaa ctgggcggag ttaggggcgg gatgggcgga gttaggggcg ggactatggt
     5281 tgctgactaa ttgagatgca tgctttgcat acttctgcct gctggggagc ctggggactt
     5341 tccacacctg gttgctgact aattgagatg catgctttgc atacttctgc ctgctgggga
     5401 gcctggggac tttccacacc ctaactgaca cacattccac agaattaatt cgcgttaaat
     5461 ttttgttaaa tcagctcatt ttttaaccaa taggccgaaa tcggcaaaat cccttataaa
     5521 tcaaaagaat agaccgagat agggttgagt gttgttccag tttggaacaa gagtccacta
     5581 ttaaagaacg tggactccaa cgtcaaaggg cgaaaaaccg tctatcaggg cgatggccca
     5641 ctacgtgaac catcacccta atcaagtttt ttggggtcga ggtgccgtaa agcactaaat
     5701 cggaacccta aagggagccc ccgatttaga gcttgacggg gaaagccggc gaacgtggcg
     5761 agaaaggaag ggaagaaagc gaaaggagcg ggcgctaggg cgctggcaag tgtagcggtc
     5821 acgctgcgcg taaccaccac acccgccgcg cttaatgcgc cgctacaggg cgcgtgggga
     5881 taccccctag agccccagct ggttctttcc gcctcagaag ccatagagcc caccgcatcc
     5941 ccagcatgcc tgctattgtc ttcccaatcc tcccccttgc tgtcctgccc caccccaccc
     6001 cccagaatag aatgacacct actcagacaa tgcgatgcaa tttcctcatt ttattaggaa
     6061 aggacagtgg gagtggcacc ttccagggtc aaggaaggca cgggggaggg gcaaacaaca
     6121 gatggctggc aactagaagg cacagtcgag gctgatcagc gggtttaaac gggccctcta
     6181 gactcgagcg gccgcctagg catttctttc ccacggttca atataatcca atcccagatg
     6241 cgcaaaaatt tcttcttcac tttctgcttt gaggaatatc ctcttggtct tgtcatataa
     6301 agcatggtta tccagaatca tcttccgctc atgtgtggca tagcgccgga ggtctctctc
     6361 aaactgccgg gagccagtcc atcccaacag ggcaaaggca cgacgctcgt aggggcacag
     6421 aactaaatcc acacggatgg ccttccaggt ctttccttcc tgccagctgg actggtcact
     6481 gtccactctt tgacgaggca atttgaaaat cagaaagcac ttttgaaaat gatccaaagc
     6541 atcaaccttc ctgctaggca acctgagctt ttcaaatgtt gactccacaa ggtcataata
     6601 taaaagtaat cccttctttt cccataagtt catcactttc tgtaaaagtt gctcttcatc
     6661 ctctgttgat cctgggctgg taattaaaaa ctctacctca tgccccatct tcttacccct
     6721 ccggaaccct cctgtcatgg tgacgaaagc atccggaaga aatgcccaga cagcctcttt
     6781 aaccagcaca ctgacggcct ctgcttctgc cctggtcaca cagctgacaa ggtcttcata
     6841 atacagaaat cctgctttct gcattcgtgt aaatttcagg cttttgtccg accttacttt
     6901 actcagagtt ctgaaaccca tcctgaacca cttctcagaa gtcttcagcc ccactccaaa
     6961 tacagaagta aagagtttga aggattgata tcgttcatca tttaacacag ctttaacttc
     7021 agaactttct ccatcttcaa taatctcctc tatgat
//
