LOCUS       Exported                7034 bp ds-DNA     circular SYN 13-JUN-2025
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7034)
  AUTHORS   Richard Davis
  TITLE     Direct Submission
  JOURNAL   Exported Tuesday, Jun 17, 2025 from SnapGene 5.0.8
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7034
                     /organism="synthetic DNA construct"
                     /mol_type="other DNA"
     protein_bind    107..128
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence 
                     of cAMP."
     promoter        143..173
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    181..197
                     /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        253..463
                     /label=SV40 promoter
                     /note="SV40 early promoter"
     promoter        559..1736
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     misc_recomb     1754..1795
                     /label=attB
                     /note="from myobacteriumphage bxb1 (Thomson et al 2012)"
     protein_bind    1808..1855
                     /label=FRT
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
                     /note="FLP-mediated recombination occurs in the 8-bp core 
                     sequence TCTAGAAA (Turan and Bode, 2011)."
     promoter        1907..3084
                     /label=EF-1-alpha promoter
                     /note="strong constitutive promoter for human elongation 
                     factor EF-1-alpha"
     regulatory      3092..3099
                     /regulatory_class="other"
                     /label=Kozak sequence
                     /note="vertebrate consensus sequence for strong initiation 
                     of translation (Kozak, 1987)"
     CDS             3107..3805
                     /codon_start=1
                     /label=mStayGold
                     /translation="MRGSDPMVSTGEELFTGVVPFKFQLKGTINGKSFTVEGEGEGNSH
                     EGSHKGKYVCTSGKLPMSWAALGTSFGYGMKYYTKYPSGLKNWFHEVMPEGFTYDRHIQ
                     YKGDGSIHAKHQHFMKNGTYHNIVEFTGQDFKENSPVLTSDMDVSLPNEVQHIPIDDGV
                     ECTVTLQYPLLSDESKCVEAYQNTIIKPLHNQPAPDVPFHWIRKQYTQSKDDTEERDHI
                     IQSETLEAHL"
     polyA_signal    3817..4041
                     /label=bGH poly(A) signal
                     /note="bovine growth hormone polyadenylation signal"
     protein_bind    4078..4125
                     /label=FRT-F3 mutant
                     /bound_moiety="FLP recombinase from the Saccharomyces 
                     cerevisiae 2u plasmid"
     rep_origin      4392..4847
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow 
                     indicates direction of (+) strand synthesis"
     promoter        5129..5233
                     /gene="bla"
                     /label=AmpR promoter
     CDS             5234..6094
                     /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"
     rep_origin      6265..6853
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
ORIGIN
        1 agcgcccaat acgcaaaccg cctctccccg cgcgttggcc gattcattaa tgcagctggc
       61 acgacaggtt tcccgactgg aaagcgggca gtgagcgcaa cgcaattaat gtgagttagc
      121 tcactcatta ggcaccccag gctttacact ttatgcttcc ggctcgtatg ttgtgtggaa
      181 ttgtgagcgg ataacaattt cacacaggaa acagctatga ccatgattac gccaagctta
      241 attccctccc cagcaggcag aagtatgcaa agcatgcatc tcaattagtc agcaaccata
      301 gtcccgcccc taactccgcc catcccgccc ctaactccgc ccagttccgc ccattctccg
      361 ccccatggct gactaatttt ttttatttat gcagaggccg aggccgcctc ggcctctgag
      421 ctattccaga agtagtgagg aggctttttt ggaggcctag gcttttgcaa aaagctttgc
      481 aaagatggat aaagttttaa acagagagga atctttgcag ctaatggacc ttctaggtct
      541 tgaaaggagt gggaattggc tccggtgccc gtcagtgggc agagcgcaca tcgcccacag
      601 tccccgagaa gttgggggga ggggtcggca attgaaccgg tgcctagaga aggtggcgcg
      661 gggtaaactg ggaaagtgat gtcgtgtact ggctccgcct ttttcccgag ggtgggggag
      721 aaccgtatat aagtgcagta gtcgccgtga acgttctttt tcgcaacggg tttgccgcca
      781 gaacacaggt aagtgccgtg tgtggttccc gcgggcctgg cctctttacg ggttatggcc
      841 cttgcgtgcc ttgaattact tccacctggc tgcagtacgt gattcttgat cccgagcttc
      901 gggttggaag tgggtgggag agttcgaggc cttgcgctta aggagcccct tcgcctcgtg
      961 cttgagttga ggcctggcct gggcgctggg gccgccgcgt gcgaatctgg tggcaccttc
     1021 gcgcctgtct cgctgctttc gataagtctc tagccattta aaatttttga tgacctgctg
     1081 cgacgctttt tttctggcaa gatagtcttg taaatgcggg ccaagatctg cacactggta
     1141 tttcggtttt tggggccgcg ggcggcgacg gggcccgtgc gtcccagcgc acatgttcgg
     1201 cgaggcgggg cctgcgagcg cggccaccga gaatcggacg ggggtagtct caagctggcc
     1261 ggcctgctct ggtgcctggc ctcgcgccgc cgtgtatcgc cccgccctgg gcggcaaggc
     1321 tggcccggtc ggcaccagtt gcgtgagcgg aaagatggcc gcttcccggc cctgctgcag
     1381 ggagctcaaa atggaggacg cggcgctcgg gagagcgggc gggtgagtca cccacacaaa
     1441 ggaaaagggc ctttccgtcc tcagccgtcg cttcatgtga ctccacggag taccgggcgc
     1501 cgtccaggca cctcgattag ttctcgagct tttggagtac gtcgtcttta ggttgggggg
     1561 aggggtttta tgcgatggag tttccccaca ctgagtgggt ggagactgaa gttaggccag
     1621 cttggcactt gatgtaattc tccttggaat ttgccctttt tgagtttgga tcttggttca
     1681 ttctcaagcc tcagacagtg gttcaaagtt tttttcttcc atttcaggtg tcgtgaggaa
     1741 ttcgccacca tggccggctt gtcgacgacg gcggactccg tcgtcaggat catccggatc
     1801 cggatccgaa gttcctattc cgaagttcct attctctaga aagtatagga acttcatgat
     1861 attcggcaag caggcatcga ctagttaatt aagtgagtgg gaattggctc cggtgcccgt
     1921 cagtgggcag agcgcacatc gcccacagtc cccgagaagt tggggggagg ggtcggcaat
     1981 tgaaccggtg cctagagaag gtggcgcggg gtaaactggg aaagtgatgt cgtgtactgg
     2041 ctccgccttt ttcccgaggg tgggggagaa ccgtatataa gtgcagtagt cgccgtgaac
     2101 gttctttttc gcaacgggtt tgccgccaga acacaggtaa gtgccgtgtg tggttcccgc
     2161 gggcctggcc tctttacggg ttatggccct tgcgtgcctt gaattacttc cacctggctg
     2221 cagtacgtga ttcttgatcc cgagcttcgg gttggaagtg ggtgggagag ttcgaggcct
     2281 tgcgcttaag gagccccttc gcctcgtgct tgagttgagg cctggcctgg gcgctggggc
     2341 cgccgcgtgc gaatctggtg gcaccttcgc gcctgtctcg ctgctttcga taagtctcta
     2401 gccatttaaa atttttgatg acctgctgcg acgctttttt tctggcaaga tagtcttgta
     2461 aatgcgggcc aagatctgca cactggtatt tcggtttttg gggccgcggg cggcgacggg
     2521 gcccgtgcgt cccagcgcac atgttcggcg aggcggggcc tgcgagcgcg gccaccgaga
     2581 atcggacggg ggtagtctca agctggccgg cctgctctgg tgcctggcct cgcgccgccg
     2641 tgtatcgccc cgccctgggc ggcaaggctg gcccggtcgg caccagttgc gtgagcggaa
     2701 agatggccgc ttcccggccc tgctgcaggg agctcaaaat ggaggacgcg gcgctcggga
     2761 gagcgggcgg gtgagtcacc cacacaaagg aaaagggcct ttccgtcctc agccgtcgct
     2821 tcatgtgact ccacggagta ccgggcgccg tccaggcacc tcgattagtt ctcgagcttt
     2881 tggagtacgt cgtctttagg ttggggggag gggttttatg cgatggagtt tccccacact
     2941 gagtgggtgg agactgaagt taggccagct tggcacttga tgtaattctc cttggaattt
     3001 gccctttttg agtttggatc ttggttcatt ctcaagcctc agacagtggt tcaaagtttt
     3061 tttcttccat ttcaggtgtc gtgaggaatt cgccaccatg gccaccatgc ggggttctga
     3121 tccgatggtg tctacaggcg aggagctgtt taccggcgtg gtgcccttca agttccagct
     3181 gaagggcacc atcaacggca agagcttcac cgtggaaggc gagggcgagg gcaatagcca
     3241 cgagggcagc cacaaaggca agtacgtgtg caccagcggc aaactgccaa tgtcttgggc
     3301 cgccctggga actagcttcg gctatggcat gaagtactac accaagtacc ccagcggcct
     3361 gaagaactgg ttccacgagg tgatgcccga gggcttcacc tacgacagac acatccagta
     3421 caagggcgac ggcagcatcc acgccaagca ccagcacttc atgaagaacg gcacctacca
     3481 caacatcgtg gagttcaccg gccaggactt caaggagaac agccccgtgc tgaccagcga
     3541 catggacgtg agcctgccca acgaggtgca gcacatcccc atagatgacg gcgtggagtg
     3601 cacagtgacc ctgcagtacc ctctgctgag cgacgagagc aagtgcgtgg aggcctacca
     3661 gaacaccatc atcaagcccc tgcacaatca gccagccccc gatgtgccat tccactggat
     3721 cagaaagcag tacacccaga gcaaggacga caccgaggag agggaccaca tcatccagag
     3781 cgagacactg gaggcccacc tgtaacaacg ttgcacctgt gccttctagt tgccagccat
     3841 ctgttgtttg cccctccccc gtgccttcct tgaccctgga aggtgccact cccactgtcc
     3901 tttcctaata aaatgaggaa attgcatcgc attgtctgag taggtgtcat tctattctgg
     3961 ggggtggggt ggggcaggac agcaaggggg aggattggga agacaatagc aggcatgctg
     4021 gggatgcggt gggctctatg gcgctcgctc gctgcaagaa ctcttcctca cggatccgaa
     4081 gttcctattc cgaagttcct attcttcaaa tagtatagga acttcagcgc tcactggccg
     4141 tcgttttaca aagggcgaat tcactggccg tcgttttaca acgtcgtgac tgggaaaacc
     4201 ctggcgttac ccaacttaat cgccttgcag cacatccccc tttcgccagc tggcgtaata
     4261 gcgaagaggc ccgcaccgat cgcccttccc aacagttgcg cagcctgaat ggcgaatggc
     4321 gcctgatgcg gtattttctc cttacgcatc tgtgcggtat ttcacaccgc atacgtcaaa
     4381 gcaaccatag tacgcgccct gtagcggcgc attaagcgcg gcgggtgtgg tggttacgcg
     4441 cagcgtgacc gctacacttg ccagcgccct agcgcccgct cctttcgctt tcttcccttc
     4501 ctttctcgcc acgttcgccg gctttccccg tcaagctcta aatcgggggc tccctttagg
     4561 gttccgattt agtgctttac ggcacctcga ccccaaaaaa cttgatttgg gtgatggttc
     4621 acgtagtggg ccatcgccct gatagacggt ttttcgccct ttgacgttgg agtccacgtt
     4681 ctttaatagt ggactcttgt tccaaactgg aacaacactc aaccctatct cgggctattc
     4741 ttttgattta taagggattt tgccgatttc ggcctattgg ttaaaaaatg agctgattta
     4801 acaaaaattt aacgcgaatt ttaacaaaat attaacgttt acaattttat ggtgcactct
     4861 cagtacaatc tgctctgatg ccgcatagtt aagccagccc cgacacccgc caacacccgc
     4921 tgacgcgccc tgacgggctt gtctgctccc ggcatccgct tacagacaag ctgtgaccgt
     4981 ctccgggagc tgcatgtgtc agaggttttc accgtcatca ccgaaacgcg cgagacgaaa
     5041 gggcctcgtg atacgcctat ttttataggt taatgtcatg ataataatgg tttcttagac
     5101 gtcaggtggc acttttcggg gaaatgtgcg cggaacccct atttgtttat ttttctaaat
     5161 acattcaaat atgtatccgc tcatgagaca ataaccctga taaatgcttc aataatattg
     5221 aaaaaggaag agtatgagta ttcaacattt ccgtgtcgcc cttattccct tttttgcggc
     5281 attttgcctt cctgtttttg ctcacccaga aacgctggtg aaagtaaaag atgctgaaga
     5341 tcagttgggt gcacgagtgg gttacatcga actggatctc aacagcggta agatccttga
     5401 gagttttcgc cccgaagaac gttttccaat gatgagcact tttaaagttc tgctatgtgg
     5461 cgcggtatta tcccgtattg acgccgggca agagcaactc ggtcgccgca tacactattc
     5521 tcagaatgac ttggttgagt actcaccagt cacagaaaag catcttacgg atggcatgac
     5581 agtaagagaa ttatgcagtg ctgccataac catgagtgat aacactgcgg ccaacttact
     5641 tctgacaacg atcggaggac cgaaggagct aaccgctttt ttgcacaaca tgggggatca
     5701 tgtaactcgc cttgatcgtt gggaaccgga gctgaatgaa gccataccaa acgacgagcg
     5761 tgacaccacg atgcctgtag caatggcaac aacgttgcgc aaactattaa ctggcgaact
     5821 acttactcta gcttcccggc aacaattaat agactggatg gaggcggata aagttgcagg
     5881 accacttctg cgctcggccc ttccggctgg ctggtttatt gctgataaat ctggagccgg
     5941 tgagcgtggg tcacgcggta tcattgcagc actggggcca gatggtaagc cctcccgtat
     6001 cgtagttatc tacacgacgg ggagtcaggc aactatggat gaacgaaata gacagatcgc
     6061 tgagataggt gcctcactga ttaagcattg gtaactgtca gaccaagttt actcatatat
     6121 actttagatt gatttaaaac ttcattttta atttaaaagg atctaggtga agatcctttt
     6181 tgataatctc atgaccaaaa tcccttaacg tgagttttcg ttccactgag cgtcagaccc
     6241 cgtagaaaag atcaaaggat cttcttgaga tccttttttt ctgcgcgtaa tctgctgctt
     6301 gcaaacaaaa aaaccaccgc taccagcggt ggtttgtttg ccggatcaag agctaccaac
     6361 tctttttccg aaggtaactg gcttcagcag agcgcagata ccaaatactg tccttctagt
     6421 gtagccgtag ttaggccacc acttcaagaa ctctgtagca ccgcctacat acctcgctct
     6481 gctaatcctg ttaccagtgg ctgctgccag tggcgataag tcgtgtctta ccgggttgga
     6541 ctcaagacga tagttaccgg ataaggcgca gcggtcgggc tgaacggggg gttcgtgcac
     6601 acagcccagc ttggagcgaa cgacctacac cgaactgaga tacctacagc gtgagctatg
     6661 agaaagcgcc acgcttcccg aagggagaaa ggcggacagg tatccggtaa gcggcagggt
     6721 cggaacagga gagcgcacga gggagcttcc agggggaaac gcctggtatc tttatagtcc
     6781 tgtcgggttt cgccacctct gacttgagcg tcgatttttg tgatgctcgt caggggggcg
     6841 gagcctatgg aaaaacgcca gcaacgcggc ctttttacgg ttcctggcct tttgctggcc
     6901 ttttgctcac atgttctttc ctgcgttatc ccctgattct gtggataacc gtattaccgc
     6961 ctttgagtga gctgataccg ctcgccgcag ccgaacgacc gagcgcagcg agtcagtgag
     7021 cgaggaagcg gaag
//
