LOCUS       Exported                6449 bp DNA     circular SYN 24-OCT-2025
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
COMMENT     SGRef: number: 1; type: "Journal Article"
FEATURES             Location/Qualifiers
     source          1..6449
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        1..105
                     /label=AmpR promoter
     rep_origin      complement(131..586)
                     /direction=LEFT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
     primer_bind     728..744
                     /label=M13 fwd
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     LTR             870..1104
                     /label=3' ITR
                     /note="piggyBac 3' inverted terminal repeat"
     promoter        1442..1645
                     /label=CMV promoter
                     /note="human cytomegalovirus (CMV) immediate early
                     promoter"
     promoter        1786..1997
                     /label=EF-1-alpha core promoter
                     /note="core promoter for human elongation factor
                     EF-1-alpha"
     LTR             2010..2278
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from human
                     T-cell leukemia virus (HTLV) type 1"
     CDS             2313..2909
                     /codon_start=1
                     /label=PuroR
                     /note="puromycin N-acetyltransferase"
                     /translation="MTEYKPTVRLATRDDVPRAVRTLAAAFADYPATRHTVDPDRHIER
                     VTELQELFLTRVGLDIGKVWVADDGAAVAVWTTPESVEAGAVFAEIGPRMAELSGSRLA
                     AQQQMEGLLAPHRPKEPAWFLATVGVSPDHQGKGLGSAVVLPGVEAAERAGVPAFLETS
                     APRNLPFYERLGFTVTADVEVPEGPRTWCMTRKPGA"
     polyA_signal    3020..3152
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     LTR             3629..3941
                     /label=5' ITR
                     /note="piggyBac 5' inverted terminal repeat"
     primer_bind     complement(4428..4444)
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     protein_bind    4452..4468
                     /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)."
     promoter        4476..4506
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    4521..4542
                     /label=CAP binding site
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     rep_origin      complement(4830..5418)
                     /direction=LEFT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     CDS             complement(5592..6449)
                     /codon_start=1
                     /label=AmpR
                     /note="beta-lactamase"
                     /translation="MSIQHFRVALIPFFAAFCLPVFAHPETLVKVKDAEDQLGARVGYI
                     ELDLNSGKILESFRPEERFPMMSTFKVLLCGAVLSRIDAGQEQLGRRIHYSQNDLVEYS
                     PVTEKHLTDGMTVRELCSAAITMSDNTAANLLLTTIGGPKELTAFLHNMGDHVTRLDRW
                     EPELNEAIPNDERDTTMPVAMATTLRKLLTGELLTLASRQQLIDWMEADKVAGPLLRSA
                     LPAGWFIADKSGAGERGSRGIIAALGPDGKPSRIVVIYTTGSQATMDERNRQIAEIGAS
                     LIKHW"
ORIGIN
        1 actcttcctt tttcaatatt attgaagcat ttatcagggt tattgtctca tgagcggata
       61 catatttgaa tgtatttaga aaaataaaca aataggggtt ccgcgcacat ttccccgaaa
      121 agtgccacct aaattgtaag cgttaatatt ttgttaaaat tcgcgttaaa tttttgttaa
      181 atcagctcat tttttaacca ataggccgaa atcggcaaaa tcccttataa atcaaaagaa
      241 tagaccgaga tagggttgag tgttgttcca gtttggaaca agagtccact attaaagaac
      301 gtggactcca acgtcaaagg gcgaaaaacc gtctatcagg gcgatggccc actacgtgaa
      361 ccatcaccct aatcaagttt tttggggtcg aggtgccgta aagcactaaa tcggaaccct
      421 aaagggagcc cccgatttag agcttgacgg ggaaagccgg cgaacgtggc gagaaaggaa
      481 gggaagaaag cgaaaggagc gggcgctagg gcgctggcaa gtgtagcggt cacgctgcgc
      541 gtaaccacca cacccgccgc gcttaatgcg ccgctacagg gcgcgtccca ttcgccattc
      601 aggctgcgca actgttggga agggcgatcg gtgcgggcct cttcgctatt acgccagctg
      661 gcgaaagggg gatgtgctgc aaggcgatta agttgggtaa cgccagggtt ttcccagtca
      721 cgacgttgta aaacgacggc cagtgagcgc gcctcgttca ttcacgtttt tgaacccgtg
      781 gaggacgggc agactcgcgg tgcaaatgtg ttttacagcg tgatggagca gatgaagatg
      841 ctcgacacgc tgcagaacac gcagctagat taaccctaga aagataatca tattgtgacg
      901 tacgttaaag ataatcatgc gtaaaattga cgcatgtgtt ttatcggtct gtatatcgag
      961 gtttatttat taatttgaat agatattaag ttttattata tttacactta catactaata
     1021 ataaattcaa caaacaattt atttatgttt atttatttat taaaaaaaaa caaaaactca
     1081 aaatttcttc tataaagtaa caaaactttt atgagggaca gccccccccc aaagccccca
     1141 gggatgtaat tacgtccctc ccccgctagg gggcagcagc gagccgcccg gggctccgct
     1201 ccggtccggc gctccccccg catccccgag ccggcagcgt gcggggacag cccgggcacg
     1261 gggaaggtgg cacgggatcg ctttcctctg aacgcttctc gctgctcttt gagcctgcag
     1321 acacctgggg ggatacgggg aaaaggcctc caaggcctac tagtattatg cccagtacat
     1381 gaccttatgg gactttccta cttggcagta catctacgta ttagtcatcg ctattaccat
     1441 ggtgatgcgg ttttggcagt acatcaatgg gcgtggatag cggtttgact cacggggatt
     1501 tccaagtctc caccccattg acgtcaatgg gagtttgttt tggcaccaaa atcaacggga
     1561 ctttccaaaa tgtcgtaaca actccgcccc attgacgcaa atgggcggta ggcgtgtacg
     1621 gtgggaggtc tatataagca gagctcgttt agtgaaccgt cagatcgcct ggagacgcca
     1681 tccacgctgt tttgacctcc atagaagatt ctagagctag cgaattcgaa tttaaatcgg
     1741 atccgcggcc gcgaaggatc tgcgatcgct ccggtgcccg tcagtgggca gagcgcacat
     1801 cgcccacagt ccccgagaag ttggggggag gggtcggcaa ttgaacgggt gcctagagaa
     1861 ggtggcgcgg ggtaaactgg gaaagtgatg tcgtgtactg gctccgcctt tttcccgagg
     1921 gtgggggaga accgtatata agtgcagtag tcgccgtgaa cgttcttttt cgcaacgggt
     1981 ttgccgccag aacacagctg aagcttcgag gggctcgcat ctctccttca cgcgcccgcc
     2041 gccctacctg aggccgccat ccacgccggt tgagtcgcgt tctgccgcct cccgcctgtg
     2101 gtgcctcctg aactgcgtcc gccgtctagg taagtttaaa gctcaggtcg agaccgggcc
     2161 tttgtccggc gctcccttgg agcctaccta gactcagccg gctctccacg ctttgcctga
     2221 ccctgcttgc tcaactctac gtctttgttt cgttttctgt tctgcgccgt tacagatcca
     2281 agctgtgacc ggcgcctacg ctagacgcca ccatgaccga gtacaagccc acggtgcgcc
     2341 tcgccacccg cgacgacgtc cccagggccg tacgcaccct cgccgccgcg ttcgccgact
     2401 accccgccac gcgccacacc gtcgatccgg accgccacat cgagcgggtc accgagctgc
     2461 aagaactctt cctcacgcgc gtcgggctcg acatcggcaa ggtgtgggtc gcggacgacg
     2521 gcgccgcggt ggcggtctgg accacgccgg agagcgtcga agcgggggcg gtgttcgccg
     2581 agatcggccc gcgcatggcc gagttgagcg gttcccggct ggccgcgcag caacagatgg
     2641 aaggcctcct ggcgccgcac cggcccaagg agcccgcgtg gttcctggcc accgtcggcg
     2701 tctcgcccga ccaccagggc aagggtctgg gcagcgccgt cgtgctcccc ggagtggagg
     2761 cggccgagcg cgccggggtg cccgccttcc tggagacctc cgcgccccgc aacctcccct
     2821 tctacgagcg gctcggcttc accgtcaccg ccgacgtcga ggtgcccgaa ggaccgcgca
     2881 cctggtgcat gacccgcaag cccggtgcct gaaatcaacc tctggattac aaaatttgtg
     2941 aaagattgac tggtattctt aactatgttg ctccttttac gctatgtgga tacgctgctt
     3001 taatgccttt gtatcagtta acttgtttat tgcagcttat aatggttaca aataaagcaa
     3061 tagcatcaca aatttcacaa ataaagcatt tttttcactg cattctagtt gtggtttgtc
     3121 caaactcatc aatgtatctt atcatgtctg gaattgactc aaatgatgtc aattagtcta
     3181 tcagaagcta tctggtctcc cttccggggg acaagacatc cctgtttaat atttaaacag
     3241 cagtgttccc aaactgggtt cttatatccc ttgctctggt caaccaggtt gcagggtttc
     3301 ctgtcctcac aggaacgaag tccctaaaga aacagtggca gccaggttta gccccggaat
     3361 tgactggatt ccttttttag ggcccattgg tatggctttt tccccgtatc cccccaggtg
     3421 tctgcaggct caaagagcag cgagaagcgt tcagaggaaa gcgatcccgt gccaccttcc
     3481 ccgtgcccgg gctgtccccg cacgctgccg gctcggggat gcggggggag cgccggaccg
     3541 gagcggagcc ccgggcggct cgctgctgcc ccctagcggg ggagggacgt aattacatcc
     3601 ctgggggctt tggggggggg ctgtccctga tatctataac aagaaaatat atatataata
     3661 agttatcacg taagtagaac atgaaataac aatataatta tcgtatgagt taaatcttaa
     3721 aagtcacgta aaagataatc atgcgtcatt ttgactcacg cggtcgttat agttcaaaat
     3781 cagtgacact taccgcattg acaagcacgc ctcacgggag ctccaagcgg cgactgagat
     3841 gtcctaaatg cacagcgacg gattcgcgct atttagaaag agagagcaat atttcaagaa
     3901 tgcatgcgtc aattttacgc agactatctt tctagggtta atctagctgc atcaggatca
     3961 tatcgtcggg tcttttttcc ggctcagtca tcgcccaagc tggcgctatc tgggcatcgg
     4021 ggaggaagaa gcccgtgcct tttcccgcga ggttgaagcg gcatggaaag agtttgccga
     4081 ggatgactgc tgctgcattg acgttgagcg aaaacgcacg tttaccatga tgattcggga
     4141 aggtgtggcc atgcacgcct ttaacggtga actgttcgtt caggccacct gggataccag
     4201 ttcgtcgcgg cttttccgga cacagttccg gatggtcagc ccgaagcgca tcagcaaccc
     4261 gaacaatacc ggcgacagcc ggaactgccg tgccggtgtg cagattaatg acagcggtgc
     4321 ggcgctggga tattacgtca gcgaggacgg gtatcctggc tggatgccgc agaaatggac
     4381 atggataccc cgtgagttac ccggcgggcg cgcttggcgt aatcatggtc atagctgttt
     4441 cctgtgtgaa attgttatcc gctcacaatt ccacacaaca tacgagccgg aagcataaag
     4501 tgtaaagcct ggggtgccta atgagtgagc taactcacat taattgcgtt gcgctcactg
     4561 cccgctttcc agtcgggaaa cctgtcgtgc cagctgcatt aatgaatcgg ccaacgcgcg
     4621 gggagaggcg gtttgcgtat tgggcgctct tccgcttcct cgctcactga ctcgctgcgc
     4681 tcggtcgttc ggctgcggcg agcggtatca gctcactcaa aggcggtaat acggttatcc
     4741 acagaatcag gggataacgc aggaaagaac atgtgagcaa aaggccagca aaaggccagg
     4801 aaccgtaaaa aggccgcgtt gctggcgttt ttccataggc tccgcccccc tgacgagcat
     4861 cacaaaaatc gacgctcaag tcagaggtgg cgaaacccga caggactata aagataccag
     4921 gcgtttcccc ctggaagctc cctcgtgcgc tctcctgttc cgaccctgcc gcttaccgga
     4981 tacctgtccg cctttctccc ttcgggaagc gtggcgcttt ctcatagctc acgctgtagg
     5041 tatctcagtt cggtgtaggt cgttcgctcc aagctgggct gtgtgcacga accccccgtt
     5101 cagcccgacc gctgcgcctt atccggtaac tatcgtcttg agtccaaccc ggtaagacac
     5161 gacttatcgc cactggcagc agccactggt aacaggatta gcagagcgag gtatgtaggc
     5221 ggtgctacag agttcttgaa gtggtggcct aactacggct acactagaag gacagtattt
     5281 ggtatctgcg ctctgctgaa gccagttacc ttcggaaaaa gagttggtag ctcttgatcc
     5341 ggcaaacaaa ccaccgctgg tagcggtggt ttttttgttt gcaagcagca gattacgcgc
     5401 agaaaaaaag gatctcaaga agatcctttg atcttttcta cggggtctga cgctcagtgg
     5461 aacgaaaact cacgttaagg gattttggtc atgagattat caaaaaggat cttcacctag
     5521 atccttttaa attaaaaatg aagttttaaa tcaatctaaa gtatatatga gtaaacttgg
     5581 tctgacagtt accaatgctt aatcagtgag gcacctatct cagcgatctg tctatttcgt
     5641 tcatccatag ttgcctgact ccccgtcgtg tagataacta cgatacggga gggcttacca
     5701 tctggcccca gtgctgcaat gataccgcga gacccacgct caccggctcc agatttatca
     5761 gcaataaacc agccagccgg aagggccgag cgcagaagtg gtcctgcaac tttatccgcc
     5821 tccatccagt ctattaattg ttgccgggaa gctagagtaa gtagttcgcc agttaatagt
     5881 ttgcgcaacg ttgttgccat tgctacaggc atcgtggtgt cacgctcgtc gtttggtatg
     5941 gcttcattca gctccggttc ccaacgatca aggcgagtta catgatcccc catgttgtgc
     6001 aaaaaagcgg ttagctcctt cggtcctccg atcgttgtca gaagtaagtt ggccgcagtg
     6061 ttatcactca tggttatggc agcactgcat aattctctta ctgtcatgcc atccgtaaga
     6121 tgcttttctg tgactggtga gtactcaacc aagtcattct gagaatagtg tatgcggcga
     6181 ccgagttgct cttgcccggc gtcaatacgg gataataccg cgccacatag cagaacttta
     6241 aaagtgctca tcattggaaa acgttcttcg gggcgaaaac tctcaaggat cttaccgctg
     6301 ttgagatcca gttcgatgta acccactcgt gcacccaact gatcttcagc atcttttact
     6361 ttcaccagcg tttctgggtg agcaaaaaca ggaaggcaaa atgccgcaaa aaagggaata
     6421 agggcgacac ggaaatgttg aatactcat
//
