LOCUS       pKG0893_pHarbor-        7256 bp    DNA     circular SYN 04-MAY-2023
DEFINITION  synthetic circular DNA
ACCESSION   .
VERSION     .
KEYWORDS    .
SOURCE      synthetic DNA construct
  ORGANISM  synthetic DNA construct
REFERENCE   1  (bases 1 to 7256)
  AUTHORS   abeitz
  TITLE     Direct Submission
  JOURNAL   Exported May 7, 2025 from SnapGene 8.0.3
            https://www.snapgene.com
FEATURES             Location/Qualifiers
     source          1..7256
                     /mol_type="other DNA"
                     /organism="synthetic DNA construct"
     promoter        42..146
                     /gene="bla"
                     /label=AmpR promoter
     CDS             147..1007
                     /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(365..384)
                     /label=oSEQ066 Amp_rev
     rep_origin      1178..1766
                     /direction=RIGHT
                     /label=ori
                     /note="high-copy-number ColE1/pMB1/pBR322/pUC origin of 
                     replication"
     primer_bind     1667..1686
                     /label=oSEQ067 pBR322-ori_fwd
     primer_bind     1920..1937
                     /label=L4440
                     /note="L4440 vector, forward primer"
     protein_bind    2054..2075
                     /label=CAP binding site
                     /bound_moiety="E. coli catabolite activator protein"
                     /note="CAP binding activates transcription in the presence
                     of cAMP."
     promoter        2090..2120
                     /label=lac promoter
                     /note="promoter for the E. coli lac operon"
     protein_bind    2128..2144
                     /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     2133..2155
                     /label=M13/pUC Reverse
                     /note="In lacZ gene"
     primer_bind     2152..2168
                     /label=M13 rev
                     /note="common sequencing primer, one of multiple similar 
                     variants"
     primer_bind     2187..2207
                     /label=T3
                     /note="T3 promoter, forward primer"
     promoter        2189..2207
                     /label=T3 promoter
                     /note="promoter for bacteriophage T3 RNA polymerase"
     promoter        2235..2461
                     /label=RSV promoter
                     /note="Rous sarcoma virus enhancer/promoter"
     LTR             2462..2642
                     /label=5' LTR (truncated)
                     /note="truncated 5' long terminal repeat (LTR) from HIV-1"
     misc_feature    2689..2814
                     /label=HIV-1 Psi
                     /note="packaging signal of human immunodeficiency virus
                     type 1"
     primer_bind     complement(2759..2779)
                     /label=oSEQ074 HIV-1_sig_rev
     misc_feature    3307..3540
                     /label=RRE
                     /note="The Rev response element (RRE) of HIV-1 allows for 
                     Rev-dependent mRNA export from the nucleus to the 
                     cytoplasm."
     misc_feature    4036..4152
                     /label=cPPT/CTS
                     /note="central polypurine tract and central termination
                     sequence of HIV-1"
     primer_bind     4085..4105
                     /label=oSEQ073 pPhage_fwd
     primer_bind     4145..4194
                     /label=LentiX1-harbor-5to3-fwd
     primer_bind     4145..4169
                     /label=LentiX1-harbor-3to5-rev
     primer_bind     4170..4202
                     /label=oHA012 blast_harbor_internal_fwd
     primer_bind     4170..4194
                     /label=LentiX1-harbor-3to5-fwd
     primer_bind     4186..4215
                     /label=oTA153 aavs_puro_R1_fwd
     primer_bind     4188..4224
                     /label=min-lenti-dest-fwd
     protein_bind    4203..4327
                     /gene="mutant version of attR"
                     /label=attR1
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(4204..4228)
                     /label=oSEQ049 attR1-rev
     primer_bind     4208..4234
                     /label=oTA170 pb_R2_rev
     primer_bind     4311..4351
                     /label=oTA155 dest-ccdB_fwd
     primer_bind     complement(4311..4351)
                     /label=oTA154 R1_d-ccdB_rev
     primer_bind     4314..4328
                     /label=Primer 2
     primer_bind     4332..4347
                     /label=oGE036 gg_entry_fwd
     promoter        join(4360..4365,4366..4383,4384..4390)
                     /label=lac UV5 promoter
                     /note="E. coli lac promoter with an ""up"" mutation"
     primer_bind     4376..4401
                     /label=Primer 1
     CDS             4444..5124
                     /codon_start=1
                     /product="chloramphenicol acetyltransferase"
                     /label=CmR
                     /note="confers resistance to chloramphenicol"
                     /translation="MEKKITGYTTVDISQWHRKEHFEAFQSVAQCTYNQTVQLDITAFL
                     KTVKKNKHKFYPAFIHILARLMNAHPEFRMAMKDGELVIWDSVHPCYTVFHEQTETFSS
                     LWSEYHDDFRQFLHIYSQDVACYGENLAYFPKGFIENMFFVSANPWVSFTSFDLNVANM
                     DNFFAPVFTMGKYYTQGDKVLMPLAIQVHHAVCDGFHVGRMLNELQQYCDEWQAGRNLE
                     DPAY"
     primer_bind     complement(4862..4893)
                     /label=oTA134 gg_internal_rev
     primer_bind     4862..4893
                     /label=oTA135 gg_internal_fwd
     CDS             5444..5749
                     /codon_start=1
                     /gene="ccdB"
                     /product="CcdB, a bacterial toxin that poisons DNA gyrase"
                     /label=ccdB
                     /note="Plasmids containing the ccdB gene cannot be 
                     propagated in standard E. coli strains."
                     /translation="MQFKVYTYKRESRYRLFVDVQSDIIDTPGRRMVIPLASARLLSDK
                     VPRELYPVVHIGDESWRMMTTDMASVPVSVIGEEVADLSHRENDIKNAINLMFWGI"
     primer_bind     5628..5646
                     /label=oSEQ012 ccdB_for
     primer_bind     5722..5791
                     /label=oTA157 single_int_R2_fwd
     primer_bind     complement(5752..5799)
                     /label=Primer 2
     primer_bind     complement(5754..5773)
                     /label=oSEQ019 SEQ_junk_check_for
     primer_bind     complement(5762..5774)
                     /label=oTA150 gg_paqc1_entry_rev
     primer_bind     complement(5785..5802)
                     /label=oTA155 dest-ccdB_fwd
     primer_bind     5785..5802
                     /label=oTA154 R1_d-ccdB_rev
     protein_bind    complement(5786..5910)
                     /gene="mutant version of attR"
                     /label=attR2
                     /bound_moiety="LR Clonase(TM)"
                     /note="recombination site for the Gateway(R) LR reaction"
     primer_bind     complement(5879..5915)
                     /label=oTA170 pb_R2_rev
     primer_bind     complement(5889..5911)
                     /label=INT-Track-pCW-rev
     primer_bind     complement(5891..5910)
                     /label=min-lenti-dest-rev
     primer_bind     complement(5895..5931)
                     /label=oHA029 PB_harbor_internal_rev
     primer_bind     complement(5908..5956)
                     /label=LentiX1-harbor-5to3-rev
     primer_bind     complement(5908..5931)
                     /label=LentiX1-harbor-3to5-rev
     misc_feature    5912
                     /label=WPRE
                     /note="woodchuck hepatitis virus posttranscriptional 
                     regulatory element"
     primer_bind     complement(5932..5956)
                     /label=LentiX1-harbor-3to5-fwd
     LTR             6009..6242
                     /label=3' LTR (Delta-U3)
                     /note="self-inactivating 3' long terminal repeat (LTR) from
                     HIV-1"
     polyA_signal    6314..6435
                     /label=SV40 poly(A) signal
                     /note="SV40 polyadenylation signal"
     rep_origin      6475..6610
                     /label=SV40 ori
                     /note="SV40 origin of replication"
     primer_bind     complement(6630..6649)
                     /label=T7
                     /note="T7 promoter, forward primer"
     promoter        complement(6631..6649)
                     /label=T7 promoter
                     /note="promoter for bacteriophage T7 RNA polymerase"
     primer_bind     complement(6659..6676)
                     /label=M13 Forward
                     /note="In lacZ gene. Also called M13-F20 or M13 (-21)
                     Forward"
     primer_bind     complement(6668..6690)
                     /label=M13/pUC Forward
                     /note="In lacZ gene"
     rep_origin      join(6817..7256,1..16)
                     /direction=RIGHT
                     /label=f1 ori
                     /note="f1 bacteriophage origin of replication; arrow
                     indicates direction of (+) strand synthesis"
ORIGIN
        1 ttaacgctta caatttaggt ggcacttttc ggggaaatgt gcgcggaacc cctatttgtt
       61 tatttttcta aatacattca aatatgtatc cgctcatgag acaataaccc tgataaatgc
      121 ttcaataata ttgaaaaagg aagagtatga gtattcaaca tttccgtgtc gcccttattc
      181 ccttttttgc ggcattttgc cttcctgttt ttgctcaccc agaaacgctg gtgaaagtaa
      241 aagatgctga agatcagttg ggtgcacgag tgggttacat cgaactggat ctcaacagcg
      301 gtaagatcct tgagagtttt cgccccgaag aacgttttcc aatgatgagc acttttaaag
      361 ttctgctatg tggcgcggta ttatcccgta ttgacgccgg gcaagagcaa ctcggtcgcc
      421 gcatacacta ttctcagaat gacttggttg agtactcacc agtcacagaa aagcatctta
      481 cggatggcat gacagtaaga gaattatgca gtgctgccat aaccatgagt gataacactg
      541 cggccaactt acttctgaca acgatcggag gaccgaagga gctaaccgct tttttgcaca
      601 acatggggga tcatgtaact cgccttgatc gttgggaacc ggagctgaat gaagccatac
      661 caaacgacga gcgtgacacc acgatgcctg tagcaatggc aacaacgttg cgcaaactat
      721 taactggcga actacttact ctagcttccc ggcaacaatt aatagactgg atggaggcgg
      781 ataaagttgc aggaccactt ctgcgctcgg cccttccggc tggctggttt attgctgata
      841 aatctggagc cggtgagcgt gggtctcgcg gtatcattgc agcactgggg ccagatggta
      901 agccctcccg tatcgtagtt atctacacga cggggagtca ggcaactatg gatgaacgaa
      961 atagacagat cgctgagata ggtgcctcac tgattaagca ttggtaactg tcagaccaag
     1021 tttactcata tatactttag attgatttaa aacttcattt ttaatttaaa aggatctagg
     1081 tgaagatcct ttttgataat ctcatgacca aaatccctta acgtgagttt tcgttccact
     1141 gagcgtcaga ccccgtagaa aagatcaaag gatcttcttg agatcctttt tttctgcgcg
     1201 taatctgctg cttgcaaaca aaaaaaccac cgctaccagc ggtggtttgt ttgccggatc
     1261 aagagctacc aactcttttt ccgaaggtaa ctggcttcag cagagcgcag ataccaaata
     1321 ctgttcttct agtgtagccg tagttaggcc accacttcaa gaactctgta gcaccgccta
     1381 catacctcgc tctgctaatc ctgttaccag tggctgctgc cagtggcgat aagtcgtgtc
     1441 ttaccgggtt ggactcaaga cgatagttac cggataaggc gcagcggtcg ggctgaacgg
     1501 ggggttcgtg cacacagccc agcttggagc gaacgaccta caccgaactg agatacctac
     1561 agcgtgagct atgagaaagc gccacgcttc ccgaagggag aaaggcggac aggtatccgg
     1621 taagcggcag ggtcggaaca ggagagcgca cgagggagct tccaggggga aacgcctggt
     1681 atctttatag tcctgtcggg tttcgccacc tctgacttga gcgtcgattt ttgtgatgct
     1741 cgtcaggggg gcggagccta tggaaaaacg ccagcaacgc ggccttttta cggttcctgg
     1801 ccttttgctg gccttttgct cacatgttct ttcctgcgtt atcccctgat tctgtggata
     1861 accgtattac cgcctttgag tgagctgata ccgctcgccg cagccgaacg accgagcgca
     1921 gcgagtcagt gagcgaggaa gcggaagagc gcccaatacg caaaccgcct ctccccgcgc
     1981 gttggccgat tcattaatgc agctggcacg acaggtttcc cgactggaaa gcgggcagtg
     2041 agcgcaacgc aattaatgtg agttagctca ctcattaggc accccaggct ttacacttta
     2101 tgcttccggc tcgtatgttg tgtggaattg tgagcggata acaatttcac acaggaaaca
     2161 gctatgacca tgattacgcc aagcgcgcaa ttaaccctca ctaaagggaa caaaagctgg
     2221 agctgcaagc ttaatgtagt cttatgcaat actcttgtag tcttgcaaca tggtaacgat
     2281 gagttagcaa catgccttac aaggagagaa aaagcaccgt gcatgccgat tggtggaagt
     2341 aaggtggtac gatcgtgcct tattaggaag gcaacagacg ggtctgacat ggattggacg
     2401 aaccactgaa ttgccgcatt gcagagatat tgtatttaag tgcctagctc gatacataaa
     2461 cgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     2521 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     2581 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     2641 cagtggcgcc cgaacaggga cttgaaagcg aaagggaaac cagaggagct ctctcgacgc
     2701 aggactcggc ttgctgaagc gcgcacggca agaggcgagg ggcggcgact ggtgagtacg
     2761 ccaaaaattt tgactagcgg aggctagaag gagagagatg ggtgcgagag cgtcagtatt
     2821 aagcggggga gaattagatc gcgatgggaa aaaattcggt taaggccagg gggaaagaaa
     2881 aaatataaat taaaacatat agtatgggca agcagggagc tagaacgatt cgcagttaat
     2941 cctggcctgt tagaaacatc agaaggctgt agacaaatac tgggacagct acaaccatcc
     3001 cttcagacag gatcagaaga acttagatca ttatataata cagtagcaac cctctattgt
     3061 gtgcatcaaa ggatagagat aaaagacacc aaggaagctt tagacaagat agaggaagag
     3121 caaaacaaaa gtaagaccac cgcacagcaa gcggccgctg atcttcagac ctggaggagg
     3181 agatatgagg gacaattgga gaagtgaatt atataaatat aaagtagtaa aaattgaacc
     3241 attaggagta gcacccacca aggcaaagag aagagtggtg cagagagaaa aaagagcagt
     3301 gggaatagga gctttgttcc ttgggttctt gggagcagca ggaagcacta tgggcgcagc
     3361 gtcaatgacg ctgacggtac aggccagaca attattgtct ggtatagtgc agcagcagaa
     3421 caatttgctg agggctattg aggcgcaaca gcatctgttg caactcacag tctggggcat
     3481 caagcagctc caggcaagaa tcctggctgt ggaaagatac ctaaaggatc aacagctcct
     3541 ggggatttgg ggttgctctg gaaaactcat ttgcaccact gctgtgcctt ggaatgctag
     3601 ttggagtaat aaatctctgg aacagatttg gaatcacacg acctggatgg agtgggacag
     3661 agaaattaac aattacacaa gcttaataca ctccttaatt gaagaatcgc aaaaccagca
     3721 agaaaagaat gaacaagaat tattggaatt agataaatgg gcaagtttgt ggaattggtt
     3781 taacataaca aattggctgt ggtatataaa attattcata atgatagtag gaggcttggt
     3841 aggtttaaga atagtttttg ctgtactttc tatagtgaat agagttaggc agggatattc
     3901 accattatcg tttcagaccc acctcccaac cccgagggga cccgacaggc ccgaaggaat
     3961 agaagaagaa ggtggagaga gagacagaga cagatccatt cgattagtga acggatctcg
     4021 acggtatcgg ttaactttta aaagaaaagg ggggattggg gggtacagtg caggggaaag
     4081 aatagtagac ataatagcaa cagacataca aactaaagaa ttacaaaaac aaattacaaa
     4141 attcaaaatt ttatcgtcca gtgtggtggc tactcatagc aggtgccact agtggagttc
     4201 cgacaagttt gtacaaaaaa gctgaacgag aaacgtaaaa tgatataaat atcaatatat
     4261 taaattagat tttgcataaa aaacagacta cataatactg taaaacacaa catatccagt
     4321 cactatggcg gattcgcggc cgcattaggc accccaggct ttacacttta tgcttccggc
     4381 tcgtataatg tgtggatttt gagttaggat ccgtcgagat tttcaggagc taaggaagct
     4441 aaaatggaga aaaaaatcac tggatatacc acagttgata tatcccaatg gcatcgtaaa
     4501 gaacattttg aggcatttca gtcagttgct caatgtacct ataaccagac cgttcagctg
     4561 gatattacgg cctttttaaa gaccgtaaag aaaaataagc acaagtttta tccggccttt
     4621 attcacattc ttgcccgcct gatgaatgct catccggaat tccgtatggc aatgaaagac
     4681 ggtgagctgg tgatatggga tagtgttcac ccttgttaca ccgttttcca tgagcaaact
     4741 gaaacgtttt catcgctctg gagtgaatac cacgacgatt tccggcagtt tctacacata
     4801 tattcgcaag atgtggcgtg ttacggtgaa aacctggcct atttccctaa agggtttatt
     4861 gagaatatgt ttttcgtttc agccaatccc tgggtgagtt tcaccagttt tgatttaaac
     4921 gtggccaata tggacaactt cttcgccccc gttttcacca tgggcaaata ttatacgcaa
     4981 ggcgacaagg tgctgatgcc gctggcgatt caggttcatc atgccgtttg tgatggcttc
     5041 catgtcggca gaatgcttaa tgaattacaa cagtactgcg atgagtggca ggcggggcgt
     5101 aatctagagg atccggctta ctaaaagcca gataacagta tgcgtatttg cgcgctgatt
     5161 tttgcggtat aagaatatat actgatatgt atacccgaag tatgtcaaaa agaggtatgc
     5221 tatgaagcag cgtattacag tgacagttga cagcgacagc tatcagttgc tcaaggcata
     5281 tatgatgtca atatctccgg tctggtaagc acaaccatgc agaatgaagc ccgtcgtctg
     5341 cgtgccgaac gctggaaagc ggaaaatcag gaagggatgg ctgaggtcgc ccggtttatt
     5401 gaaatgaacg gctcttttgc tgacgagaac aggggctggt gaaatgcagt ttaaggttta
     5461 cacctataaa agagagagcc gttatcgtct gtttgtggat gtacagagtg atattattga
     5521 cacgcccggg cgacggatgg tgatccccct ggccagtgca cgtctgctgt cagataaagt
     5581 cccccgtgaa ctttacccgg tggtgcatat cggggatgaa agctggcgca tgatgaccac
     5641 cgatatggcc agtgtgccgg tttccgttat cggggaagaa gtggctgatc tcagccaccg
     5701 cgaaaatgac atcaaaaacg ccattaacct gatgttctgg ggaatataaa tgtcaggctc
     5761 ccttatacac agcctgcagg tcgaccatag tgactggata tgttgtgttt tacagtatta
     5821 tgtagtctgt tttttatgca aaatctaatt taatatattg atatttatat cattttacgt
     5881 ttctcgttca gctttcttgt acaaagtggt cacctgctat gcgctgatat agggcccggt
     5941 acctttaaga ccaatgactt acaaggcagc tgtagatctt agccactttt taaaagaaaa
     6001 ggggggactg gaagggctag ctcactccca acgaagacaa gatctgcttt ttgcttgtac
     6061 tgggtctctc tggttagacc agatctgagc ctgggagctc tctggctaac tagggaaccc
     6121 actgcttaag cctcaataaa gcttgccttg agtgcttcaa gtagtgtgtg cccgtctgtt
     6181 gtgtgactct ggtaactaga gatccctcag acccttttag tcagtgtgga aaatctctag
     6241 cagtagtagt tcatgtcatc ttattattca gtatttataa cttgcaaaga aatgaatatc
     6301 agagagtgag aggaacttgt ttattgcagc ttataatggt tacaaataaa gcaatagcat
     6361 cacaaatttc acaaataaag catttttttc actgcattct agttgtggtt tgtccaaact
     6421 catcaatgta tcttatcatg tctggctcta gctatcccgc ccctaactcc gcccatcccg
     6481 cccctaactc cgcccagttc cgcccattct ccgccccatg gctgactaat tttttttatt
     6541 tatgcagagg ccgaggccgc ctcggcctct gagctattcc agaagtagtg aggaggcttt
     6601 tttggaggcc tagggacgta cccaattcgc cctatagtga gtcgtattac gcgcgctcac
     6661 tggccgtcgt tttacaacgt cgtgactggg aaaaccctgg cgttacccaa cttaatcgcc
     6721 ttgcagcaca tccccctttc gccagctggc gtaatagcga agaggcccgc accgatcgcc
     6781 cttcccaaca gttgcgcagc ctgaatggcg aatgggacgc gccctgtagc ggcgcattaa
     6841 gcgcggcggg tgtggtggtt acgcgcagcg tgaccgctac acttgccagc gccctagcgc
     6901 ccgctccttt cgctttcttc ccttcctttc tcgccacgtt cgccggcttt ccccgtcaag
     6961 ctctaaatcg ggggctccct ttagggttcc gatttagtgc tttacggcac ctcgacccca
     7021 aaaaacttga ttagggtgat ggttcacgta gtgggccatc gccctgatag acggtttttc
     7081 gccctttgac gttggagtcc acgttcttta atagtggact cttgttccaa actggaacaa
     7141 cactcaaccc tatctcggtc tattcttttg atttataagg gattttgccg atttcggcct
     7201 attggttaaa aaatgagctg atttaacaaa aatttaacgc gaattttaac aaaata
//
