LOCUS       Exported                6449 bp DNA     circular SYN 09-AUG-2024
DEFINITION  synthetic circular DNA.
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 6449)
  TITLE     Direct Submission
REFERENCE   2  (bases 1 to 6449)
  AUTHORS   .
  TITLE     Direct Submission
  JOURNAL   Exported Aug 9, 2024 from SnapGene 6.0.2
            https://www.snapgene.com
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     CDS             1..858
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
     rep_origin      1032..1620
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     protein_bind    1908..1929
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        1944..1974
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    1982..1998
                     /label=lac operator
                     /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     2006..2022
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             2509..2821
                     /label=5' ITR
                     /note="piggyBac 5' inverted terminal repeat"
     polyA_signal    3298..3430
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     CDS             complement(3541..4137)
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     LTR             4172..4440
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     promoter        4453..4664
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     promoter        4805..5008
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     LTR             complement(5346..5580)
                     /label=3' ITR
                     /note="piggyBac 3' inverted terminal repeat"
     primer_bind     complement(5706..5722)
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     rep_origin      5864..6319
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     promoter        6345..6449
                     /label=AmpR promoter
ORIGIN
        1 atgagtattc aacatttccg tgtcgccctt attccctttt ttgcggcatt ttgccttcct
       61 gtttttgctc acccagaaac gctggtgaaa gtaaaagatg ctgaagatca gttgggtgca
      121 cgagtgggtt acatcgaact ggatctcaac agcggtaaga tccttgagag ttttcgcccc
      181 gaagaacgtt ttccaatgat gagcactttt aaagttctgc tatgtggcgc ggtattatcc
      241 cgtattgacg ccgggcaaga gcaactcggt cgccgcatac actattctca gaatgacttg
      301 gttgagtact caccagtcac agaaaagcat cttacggatg gcatgacagt aagagaatta
      361 tgcagtgctg ccataaccat gagtgataac actgcggcca acttacttct gacaacgatc
      421 ggaggaccga aggagctaac cgcttttttg cacaacatgg gggatcatgt aactcgcctt
      481 gatcgttggg aaccggagct gaatgaagcc ataccaaacg acgagcgtga caccacgatg
      541 cctgtagcaa tggcaacaac gttgcgcaaa ctattaactg gcgaactact tactctagct
      601 tcccggcaac aattaataga ctggatggag gcggataaag ttgcaggacc acttctgcgc
      661 tcggcccttc cggctggctg gtttattgct gataaatctg gagccggtga gcgtgggtct
      721 cgcggtatca ttgcagcact ggggccagat ggtaagccct cccgtatcgt agttatctac
      781 acgacgggga gtcaggcaac tatggatgaa cgaaatagac agatcgctga gataggtgcc
      841 tcactgatta agcattggta actgtcagac caagtttact catatatact ttagattgat
      901 ttaaaacttc atttttaatt taaaaggatc taggtgaaga tcctttttga taatctcatg
      961 accaaaatcc cttaacgtga gttttcgttc cactgagcgt cagaccccgt agaaaagatc
     1021 aaaggatctt cttgagatcc tttttttctg cgcgtaatct gctgcttgca aacaaaaaaa
     1081 ccaccgctac cagcggtggt ttgtttgccg gatcaagagc taccaactct ttttccgaag
     1141 gtaactggct tcagcagagc gcagatacca aatactgtcc ttctagtgta gccgtagtta
     1201 ggccaccact tcaagaactc tgtagcaccg cctacatacc tcgctctgct aatcctgtta
     1261 ccagtggctg ctgccagtgg cgataagtcg tgtcttaccg ggttggactc aagacgatag
     1321 ttaccggata aggcgcagcg gtcgggctga acggggggtt cgtgcacaca gcccagcttg
     1381 gagcgaacga cctacaccga actgagatac ctacagcgtg agctatgaga aagcgccacg
     1441 cttcccgaag ggagaaaggc ggacaggtat ccggtaagcg gcagggtcgg aacaggagag
     1501 cgcacgaggg agcttccagg gggaaacgcc tggtatcttt atagtcctgt cgggtttcgc
     1561 cacctctgac ttgagcgtcg atttttgtga tgctcgtcag gggggcggag cctatggaaa
     1621 aacgccagca acgcggcctt tttacggttc ctggcctttt gctggccttt tgctcacatg
     1681 ttctttcctg cgttatcccc tgattctgtg gataaccgta ttaccgcctt tgagtgagct
     1741 gataccgctc gccgcagccg aacgaccgag cgcagcgagt cagtgagcga ggaagcggaa
     1801 gagcgcccaa tacgcaaacc gcctctcccc gcgcgttggc cgattcatta atgcagctgg
     1861 cacgacaggt ttcccgactg gaaagcgggc agtgagcgca acgcaattaa tgtgagttag
     1921 ctcactcatt aggcacccca ggctttacac tttatgcttc cggctcgtat gttgtgtgga
     1981 attgtgagcg gataacaatt tcacacagga aacagctatg accatgatta cgccaagcgc
     2041 gcccgccggg taactcacgg ggtatccatg tccatttctg cggcatccag ccaggatacc
     2101 cgtcctcgct gacgtaatat cccagcgccg caccgctgtc attaatctgc acaccggcac
     2161 ggcagttccg gctgtcgccg gtattgttcg ggttgctgat gcgcttcggg ctgaccatcc
     2221 ggaactgtgt ccggaaaagc cgcgacgaac tggtatccca ggtggcctga acgaacagtt
     2281 caccgttaaa ggcgtgcatg gccacacctt cccgaatcat catggtaaac gtgcgttttc
     2341 gctcaacgtc aatgcagcag cagtcatcct cggcaaactc tttccatgcc gcttcaacct
     2401 cgcgggaaaa ggcacgggct tcttcctccc cgatgcccag atagcgccag cttgggcgat
     2461 gactgagccg gaaaaaagac ccgacgatat gatcctgatg cagctagatt aaccctagaa
     2521 agatagtctg cgtaaaattg acgcatgcat tcttgaaata ttgctctctc tttctaaata
     2581 gcgcgaatcc gtcgctgtgc atttaggaca tctcagtcgc cgcttggagc tcccgtgagg
     2641 cgtgcttgtc aatgcggtaa gtgtcactga ttttgaacta taacgaccgc gtgagtcaaa
     2701 atgacgcatg attatctttt acgtgacttt taagatttaa ctcatacgat aattatattg
     2761 ttatttcatg ttctacttac gtgataactt attatatata tattttcttg ttatagatat
     2821 cagggacagc ccccccccaa agcccccagg gatgtaatta cgtccctccc ccgctagggg
     2881 gcagcagcga gccgcccggg gctccgctcc ggtccggcgc tccccccgca tccccgagcc
     2941 ggcagcgtgc ggggacagcc cgggcacggg gaaggtggca cgggatcgct ttcctctgaa
     3001 cgcttctcgc tgctctttga gcctgcagac acctgggggg atacggggaa aaagccatac
     3061 caatgggccc taaaaaagga atccagtcaa ttccggggct aaacctggct gccactgttt
     3121 ctttagggac ttcgttcctg tgaggacagg aaaccctgca acctggttga ccagagcaag
     3181 ggatataaga acccagtttg ggaacactgc tgtttaaata ttaaacaggg atgtcttgtc
     3241 ccccggaagg gagaccagat agcttctgat agactaattg acatcatttg agtcaattcc
     3301 agacatgata agatacattg atgagtttgg acaaaccaca actagaatgc agtgaaaaaa
     3361 atgctttatt tgtgaaattt gtgatgctat tgctttattt gtaaccatta taagctgcaa
     3421 taaacaagtt aactgataca aaggcattaa agcagcgtat ccacatagcg taaaaggagc
     3481 aacatagtta agaataccag tcaatctttc acaaattttg taatccagag gttgatttca
     3541 ggcaccgggc ttgcgggtca tgcaccaggt gcgcggtcct tcgggcacct cgacgtcggc
     3601 ggtgacggtg aagccgagcc gctcgtagaa ggggaggttg cggggcgcgg aggtctccag
     3661 gaaggcgggc accccggcgc gctcggccgc ctccactccg gggagcacga cggcgctgcc
     3721 cagacccttg ccctggtggt cgggcgagac gccgacggtg gccaggaacc acgcgggctc
     3781 cttgggccgg tgcggcgcca ggaggccttc catctgttgc tgcgcggcca gccgggaacc
     3841 gctcaactcg gccatgcgcg ggccgatctc ggcgaacacc gcccccgctt cgacgctctc
     3901 cggcgtggtc cagaccgcca ccgcggcgcc gtcgtccgcg acccacacct tgccgatgtc
     3961 gagcccgacg cgcgtgagga agagttcttg cagctcggtg acccgctcga tgtggcggtc
     4021 cggatcgacg gtgtggcgcg tggcggggta gtcggcgaac gcggcggcga gggtgcgtac
     4081 ggccctgggg acgtcgtcgc gggtggcgag gcgcaccgtg ggcttgtact cggtcatggt
     4141 ggcgtctagc gtaggcgccg gtcacagctt ggatctgtaa cggcgcagaa cagaaaacga
     4201 aacaaagacg tagagttgag caagcagggt caggcaaagc gtggagagcc ggctgagtct
     4261 aggtaggctc caagggagcg ccggacaaag gcccggtctc gacctgagct ttaaacttac
     4321 ctagacggcg gacgcagttc aggaggcacc acaggcggga ggcggcagaa cgcgactcaa
     4381 ccggcgtgga tggcggcctc aggtagggcg gcgggcgcgt gaaggagaga tgcgagcccc
     4441 tcgaagcttc agctgtgttc tggcggcaaa cccgttgcga aaaagaacgt tcacggcgac
     4501 tactgcactt atatacggtt ctcccccacc ctcgggaaaa aggcggagcc agtacacgac
     4561 atcactttcc cagtttaccc cgcgccacct tctctaggca cccgttcaat tgccgacccc
     4621 tccccccaac ttctcgggga ctgtgggcga tgtgcgctct gcccactgac gggcaccgga
     4681 gcgatcgcag atccttcgcg gccgcggatc cgatttaaat tcgaattcgc tagctctaga
     4741 atcttctatg gaggtcaaaa cagcgtggat ggcgtctcca ggcgatctga cggttcacta
     4801 aacgagctct gcttatatag acctcccacc gtacacgcct accgcccatt tgcgtcaatg
     4861 gggcggagtt gttacgacat tttggaaagt cccgttgatt ttggtgccaa aacaaactcc
     4921 cattgacgtc aatggggtgg agacttggaa atccccgtga gtcaaaccgc tatccacgcc
     4981 cattgatgta ctgccaaaac cgcatcacca tggtaatagc gatgactaat acgtagatgt
     5041 actgccaagt aggaaagtcc cataaggtca tgtactgggc ataatactag taggccttgg
     5101 aggccttttc cccgtatccc cccaggtgtc tgcaggctca aagagcagcg agaagcgttc
     5161 agaggaaagc gatcccgtgc caccttcccc gtgcccgggc tgtccccgca cgctgccggc
     5221 tcggggatgc ggggggagcg ccggaccgga gcggagcccc gggcggctcg ctgctgcccc
     5281 ctagcggggg agggacgtaa ttacatccct gggggctttg ggggggggct gtccctcata
     5341 aaagttttgt tactttatag aagaaatttt gagtttttgt ttttttttaa taaataaata
     5401 aacataaata aattgtttgt tgaatttatt attagtatgt aagtgtaaat ataataaaac
     5461 ttaatatcta ttcaaattaa taaataaacc tcgatataca gaccgataaa acacatgcgt
     5521 caattttacg catgattatc tttaacgtac gtcacaatat gattatcttt ctagggttaa
     5581 tctagctgcg tgttctgcag cgtgtcgagc atcttcatct gctccatcac gctgtaaaac
     5641 acatttgcac cgcgagtctg cccgtcctcc acgggttcaa aaacgtgaat gaacgaggcg
     5701 cgctcactgg ccgtcgtttt acaacgtcgt gactgggaaa accctggcgt tacccaactt
     5761 aatcgccttg cagcacatcc ccctttcgcc agctggcgta atagcgaaga ggcccgcacc
     5821 gatcgccctt cccaacagtt gcgcagcctg aatggcgaat gggacgcgcc ctgtagcggc
     5881 gcattaagcg cggcgggtgt ggtggttacg cgcagcgtga ccgctacact tgccagcgcc
     5941 ctagcgcccg ctcctttcgc tttcttccct tcctttctcg ccacgttcgc cggctttccc
     6001 cgtcaagctc taaatcgggg gctcccttta gggttccgat ttagtgcttt acggcacctc
     6061 gaccccaaaa aacttgatta gggtgatggt tcacgtagtg ggccatcgcc ctgatagacg
     6121 gtttttcgcc ctttgacgtt ggagtccacg ttctttaata gtggactctt gttccaaact
     6181 ggaacaacac tcaaccctat ctcggtctat tcttttgatt tataagggat tttgccgatt
     6241 tcggcctatt ggttaaaaaa tgagctgatt taacaaaaat ttaacgcgaa ttttaacaaa
     6301 atattaacgc ttacaattta ggtggcactt ttcggggaaa tgtgcgcgga acccctattt
     6361 gtttattttt ctaaatacat tcaaatatgt atccgctcat gagacaataa ccctgataaa
     6421 tgcttcaata atattgaaaa aggaagagt
//
